Journal Club Presentation

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3. https://www.cdc.gov/drugresistance/threat-report-2013/ https://www.cdc.gov/foodsafety/challenges/from-farm-to-table.html
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4. Colistin Case China
• Last resort drug
• Damages liver and kidney
• Used recently to treat ventilator-
associated pneumonia (VAP) and
bacteremia caused by MDR bacteria, such
as P. aeruginosa, K. pneumoniae and A.
baumannii.
100,000 flights daily
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The Antibiotic Apocalypse Explained- Kurzgesagt – In a Nutshell

5. The Review on Antimicrobial Resistance- by Jim O’Neill
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6. http://www.biomedcentral.com/content/figures/1741-7007-8-123-1-l.jpg
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(Penicillin binding proteins)

7. Gerard D Wright - http://www.biomedcentral.com/content/figures/1741-7007-8-123-1-l.jpg
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8. • A set of human technologies are hotspots and mixing vessels of
pollutants, which force bacteria to maintain ARGs in water.
• ARGs can be dispersed between
– sylvatic reservoirs
– domestic animals
– cultivated vegetables
– and human populations
(Arnold et al., 2016)
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9. • Technologies responsible for outstanding gains in human survival
are now being implicated in the multi-resistance health crisis:
– sewerage-sanitation
– antibiotics
– animal husbandry
– and crop production
• Wastewater treatment plants (WWTPs) are a chief release point of
antibiotics and ARGs.
• Humans and domestic animals are amplifying hosts for a
pathogen; are often co-infected by several pathogens, and add to
waterborne ARG transfers.
(Ferriman, 2007; Laxminarayan et al., 2013; Roca et al.,
2015)
(Giannakis et al., 2016; Michael et al., 2013)
(De las Heras et al., 2016)
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10. (Marina et al. 2018)
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11. • ARGs are more difficult to destroy than pathogens.
– found in plasmids which commonly exist in water and facilitate the
transmission of ARGs.
Antibacterial, Metal and ARGs (AMA) pollution should be treated in
situ (by industries, hospitals and households) to separate AMA
pollutants, unlike WWTP that mix and disperse pollutants.
(Giannakis et al., 2016)
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12. Combination of natural bacterial
degradation capabilities can fulfil
the depuration functions in
wastewaters polluted with
antibacterials, metals and DNA
(ARGs and potential pathogens).
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To measure the efficiency of unit
operations in a treatment train
capable of remediating water
polluted with antibacterials,
metals and DNA.
1. Aerobic treatment of
antibacterial pollution
2. Aerobic treatment of metal
pollution
3. Anaerobic treatment of
antibacterial pollution
4. Thermo-pressure degradation
of DNA

13. Four unit operations (Fig. 2) were tested
individually, to assess in the laboratory the
viability of coupling and scaling them up in
a prototype.
To determine a better sequence of
operations and size of the apparatus.
Factors considered:
– efficiency and rate of biodegradation
– initial pollutant concentration
– and volumetric flows
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14. Aerobic catabolic activity of the aromatic-ring antibacterials 14C-labeled Sulfamethazine (SMZ) and 14C-labelled
Erythromycin (ERY) was determined by respirometry assays.
These faster catabolism of aerobic antibacterial-resistant bacteria was used to mitigate antibacterial peaks that jeopardize
optimal operation of AD.
Four bacteria that showed multidrug resistance (ERY, SMZ, Chloramphenicol (CHLO), Tetracycline (TET) and
Kanamycin (KAN)), high MICs and grew in presence of the antibacterials, were selected and pooled into a
consortium.
Bacterial 16S rRNA genes from isolated bacteria were amplified by PCR for identification, and known bacterial
pathogens were excluded.
The bacteria were isolated from soil and/or manure from pigs treated with antibacterials.
(Islas-Espinoza et al., 14

15. Cellular sorption or accumulation of CrVI, Pb and Cd in the bacteria were ascertained via freezing-thawing thermal
shock to induce cell lysis, release and resuspend the metals in water.
Growth of the bacteria in the presence of metals was measured via optical density spectrophotometry (OD600).
Total Cr, Cd and Pb were determined using atomic absorption, and UV-Vis spectrophotometry for CrVI
Bioreduction of CrVI to CrIII was assessed as well as biotransformations of Pb, Cd and the mixture of CrVI, Pb and
Cd.
Bioremediation of metal mixtures in chromium-plating wastewater prior to anaerobic digestion (AD) was conducted
with the bacteria Brevundimonas diminuta, which was isolated from a Cr plating workshop.
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16. After acclimation, SMX and ERY were spiked at increasing concentrations in the influent until CH4 production
collapsed at which point antibacterial spiking stopped to observe possible recovery
The control ASBR was fed synthetic wastewater, another ASBR was spiked with SMX and the other ASBR with
ERY+SMX
A synthetic substrate was used to resemble wastewater from a pharmaceutical industry.
Three anaerobic sequencing batch reactors (ASBRs) were inoculated with anaerobic granular sludge.
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17. • During anaerobiosis, mRNA copies of genes encoding
– formyltetrahydrofolate synthetase (FTHFS),
– methyl-coenzyme M reductase (mcrA) and
– acetyl-coA synthetase (ACAS)
were targeted to measure enzymatic activity inhibition in
homoacetogens, methanogens and acetoclastic methanogens,
respectively.
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18. The breaking of the covalent bonds of DNA was determined by gel electrophoresis using 1% agarose, for 60 min at
70 V.
Additionally, E. coli colonies (resistant to 100mg/L ERY, SMX and CHLO, separately) were tested for inactivation in
the same tubes.
Temperatures tested were 20, 40, 60, 80, 90, 100 and 150°C and pressures were observed in the 1.4 - 16 kPa gauge
pressure range.
Tick and chicken kidney DNA were extracted by phenol-chloroform extraction and ethanol precipitation.
Escherichia coli DNA was extracted with an UltraClean Water DNA kit
(Girish et al., 2005)
(Karni et al., 2013)
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24. Initial concentrations in aerobic concentrations were one order of
magnitude higher, justifying them as pre-treatments to the anaerobic
digestion (AD).
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25. Result Table
Treatment Type Pollutant Treatment Duration (Days) Treatment Efficiency
Aerobic
Cr(VI) reduction
8 25.28-71.88%
Cd and Pb removal
14C-Erythromycin (ERY) 20 34.77%
14C-Sulfamethazine (SMZ) 20 7.83%
Chloramphenicol (CHLO) 10 31.00%
Tetracycline (TET) 10 20.00%
Anaerobic Digestion
(AD)
Erythromycin (ERY)
60 65.0-73.0%
Sulfamethoxazole (SMX)
Thermo-pressure
degradation
DNA (Antibacterial Resistance
Genes)
1/6 100.0%
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27. This was the first proof-of-concept of a sustainable bioremediation train
aimed at modifying wastewater technologies implicated in multi-
resistance.
Bacterial resistance was harnessed for aromatic ring antibacterial and
metal bioremediation, water organic pollution, and production of
biomethane.
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28. This was the first proof-of-concept of a sustainable bioremediation train aimed at modifying wastewater technologies implicated in
multi-resistance.
Bacterial resistance was harnessed for aromatic ring antibacterial and metal bioremediation, water organic pollution, and
production of biomethane.
Biodegradation was initiated and enhanced by biodiverse communities.
ARGs must be treated in a controlled environment to avoid dispersion.
A combination of bacterial capabilities may be organized in specifically
targeted pre-treatments to smooth out pollution peaks in the influent of the
treatment.
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29. This was the first proof-of-concept of a sustainable bioremediation train aimed at modifying wastewater technologies implicated in multi-
resistance.
Bacterial resistance was harnessed for aromatic ring antibacterial and metal bioremediation, water organic pollution, and production of
biomethane.
Biodegradation was initiated and enhanced by biodiverse communities. ARGs must be treated in a controlled environment to avoid
dispersion.
A combination of bacterial capabilities may be organized in specifically targeted pre-treatments to smooth out pollution peaks in
the influent of the treatment.
The disinfection post-treatment avoids releases of the pollutants.
The sustainability of the system hinges on the use of wastewater organic matter
as energy and nutrient source for the biodegraders, and for the production of an
energy carrier (CH4) useful in disinfection.
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30. This was the first proof-of-concept of a sustainable bioremediation train aimed at modifying wastewater technologies implicated in multi-
resistance.
Bacterial resistance was harnessed for aromatic ring antibacterial and metal bioremediation, water organic pollution, and production of
biomethane.
Biodegradation was initiated and enhanced by biodiverse communities. ARGs must be treated in a controlled environment to avoid
dispersion.
A combination of bacterial capabilities may be organized in specifically targeted pre-treatments to smooth out pollution peaks in
the influent of the treatment.
The disinfection post-treatment avoids releases of the pollutants.
The sustainability of the system hinges on the use of wastewater organic matter as energy and nutrient source for the
biodegraders, and for the production of an energy carrier (CH4) useful in disinfection.
Biomedical advances such as discovery of new antibacterials, and policies such as
reduced antibacterial use, need to be complemented by sustainable technologies
that prevent resistance genes, antibacterial and metal accumulations in the
environment.
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31. This was the first proof-of-concept of a sustainable bioremediation train aimed at modifying wastewater technologies implicated in multi-
resistance.
Bacterial resistance was harnessed for aromatic ring antibacterial and metal bioremediation, water organic pollution, and production of
biomethane.
Biodegradation was initiated and enhanced by biodiverse communities. ARGs must be treated in a controlled environment to avoid
dispersion.
A combination of bacterial capabilities may be organized in specifically targeted pre-treatments to smooth out pollution peaks in
the influent of the treatment.
The disinfection post-treatment avoids releases of the pollutants.
The sustainability of the system hinges on the use of wastewater organic matter as energy and nutrient source for the biodegraders, and
for the production of an energy carrier (CH4) useful in disinfection.
Biomedical advances such as discovery of new antibacterials, and policies such as reduced antibacterial use, need to be complemented by
sustainable technologies that prevent resistance genes, antibacterial and metal accumulations in the environment.
31

32. • "What is Drug Resistance?". www.niaid.nih.gov. Retrieved 2019-07-26.
• Arnold, K.E., Williams, N.J., Bennett, M., 2016. “Disperse abroad in the land”: the role of wildlife in the dissemination of antimicrobial
resistance. Biol. Lett. 12, 1e8. https://doi.org/dx.doi.org/10.1098/rsbl.2016.0137.
• Aydin, S., Ince, B., Cetecioglu, Z., Arikan, O., Ozbayram, E.G., Shahi, A., Ince, O., 2015a. Combined effect of erythromycin, tetracycline
and sulfamethoxazole on performance of anaerobic sequencing batch reactors. Bioresour. Technol. 186, 207e214.
• Aydin, S., Ince, B., Ince, O., 2015b. Development of antibiotic resistance genes in microbial communities during long-term operation of
anaerobic reactors in the treatment of pharmaceutical wastewater. Water Res. 83, 337e344.
• De las Heras, A., Islas-Espinoza, M., Amaya-Chavez, A., 2016. Pollution: the pathogenic and xenobiotic exposome of humans and the
need for technological change. Encycl. Environ. Manag. 1e10. https://doi.org/10.1081/E-EEM- 120052926.
• Ferriman, A., 2007. BMJ readers choose sanitation as greatest medical advance since 1840. BMJ 334, 111.
• Gerard D Wright - http://www.biomedcentral.com/content/figures/1741-7007-8-123-1-l.jpg
• Giannakis, S., Inmaculada, M., Lopez, P., Spuhler, D., Antonio, J., Perez, S., Fernandez, P., 2016. Solar disinfection is an augmentable, in
situ-generated photo-Fenton reaction - Part 2: a review of the applications for drinking water and wastewater disinfection. Appl. Catal.
B Environ. 198, 431e446. https://doi.org/10.1016/j.apcatb.2016.06.007.
• Giannakis, S., Inmaculada, M., Lopez, P., Spuhler, D., Antonio, J., Perez, S., Fernandez, P., 2016. Solar disinfection is an augmentable, in
situ-generated photo-Fenton reaction d Part 2: a review of the applications for drinking water and wastewater disinfection. Appl. Catal.
B Environ. 198, 431e446. https://doi.org/10.1016/j.apcatb.2016.06.007.
• Girish, P., Anjaneyulu, A., Viswas, K., Shivakumar, B., Anand, M., Patel, M., Sharma, B., 2005. Meat species identification by polymerase
chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Sci. 70, 107e112.
https://doi.org/10.1016/j.meatsci.2004.12.004.
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33. • http://www.biomedcentral.com/content/figures/1741-7007-8-123-1-l.jpg
• https://www.cdc.gov/foodsafety/challenges/from-farm-to-table.html
• Infographic from the CDC Threat Report 2013, Antibiotic resistance threats in the United States, 2013
(https://www.cdc.gov/drugresistance/threat-report-2013/)
• Islas-Espinoza, M., Reid, B.J.,Wexler, M., Bond, P.L., 2012. Soil bacterial consortia and previous exposure enhance the biodegradation of
sulfonamides from pig manure. Microb. Ecol. 64, 140e151.
• Karni, M., Zidon, D., Polak, P., Zalevsky, Z., Shefi, O., 2013. Thermal degradation of DNA. DNA Cell Biol. 32, 1e4.
https://doi.org/10.1089/dna.2013.2056.
• Laxminarayan, R., Duse, A., Wattal, C., Zaidi, A.K.M., Wertheim, H.F.L., Sumpradit, N., Peralta, A.Q., 2013. The Lancet infectious diseases
commission antibiotic resistance - the need for global solutions, 3099 (13).
• Michael, I., Rizzo, L., Mcardell, C.S., Manaia, C.M., Merlin, C., Schwartz, T., Fattakassinos, D., 2013. Urban wastewater treatment plants
as hotspots for the release of antibiotics in the environment: a review. Water Res. 47 (3), 957e995.
https://doi.org/10.1016/j.watres.2012.11.027.
• Roca, I., Akova, M., Baquero, F., Carlet, J., Cavaleri, M., Coenen, S., Heure, O.E., 2015. The global threat of antimicrobial resistance:
science for intervention. New Microbes New Infect. 6, 22e29. https://doi.org/10.1016/j.nmni.2015.02.007.
• The Antibiotic Apocalypse Explained- Kurzgesagt – In a Nutshell
• The Review on Antimicrobial Resistance, Chaired by Jim O’Neill
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