Emerging Contaminants

Emerging Contaminants
• Pharmaceutical drugs include analgesics, antibiotics, contraceptives,
lipid regulators, beta blockers and steroidal hormones…

• These organic compounds are part of the emerging organic pollutants

that are now being detected in water systems, probably as a result of
their increased use and/or the improvements that have taken place in
analytical techniques…
 Pharmaceutical Contaminants
• Criteria that usually influence the selection of drugs following
fundamental points:

Prescription volumes
The toxicity of parent compounds, as well as their metabolites and
transformation products
Adverse health effects on both humans and animals (such as
carcinogenicity, mutagenicity and endocrine disruption)
 Pharmaceutical Contaminants
• Criteria that usually influence the selection of drugs following
fundamental points:

The stability or persistence of the drugs in the environment

The removal efficiencies of the drugs when treated using conventional
water treatment systems
Degradation and photolysis
Pharmaceutical Contaminants
• According to this priority list, the most prevalent groups of
pharmaceuticals from both the public and private health sector are
analgesics, hypertension drugs, antihistamines, vitamins, ARVs,
NSAIDs, antidiabetics and antibiotics…

• Hence, the pharmaceutical residues that have been detected in South

African water systems also fall within this list
Pharmaceutical Contaminants
• In addition to the criteria listed above, a suggested protocol for
monitoring of organic pollutants in drinking water is as follows:

The potential of finding the compound in drinking water

The availability of standards and guidelines for regulation
The ease of monitoring in the drinking water value chain
Pharmaceutical Contaminants
• In addition to the criteria listed above, a suggested protocol for
monitoring of organic pollutants in drinking water is as follows:

The potential of the contaminant to cause aesthetic water quality

problems
The potential to increase customer perception risk
Pharmaceutical Contaminants

• After use by humans and animals, many drugs are excreted without
being metabolised by the patients & consequently enter wastewater
through the sewage systems either in their parent or their
metabolised form…
Pharmaceutical Contaminants

• Emerging contaminants enter the environment, specifically water,

through a variety of pathways that can be categorised as point source
(municipal sewage, industrial wastewaters and landfill) & non-point
source (agricultural run-off, wash-off from roadways and
underground contamination)…
Pharmaceutical Contaminants

• Most of the countries WWTPs are not designed to remove most of

the emerging contaminants…

• Hence, a high portion of emerging compounds & their metabolites

can escape & enter the environment via sewage effluents…
Agricultural Contaminants
• In terms of agricultural systems, there are a number of EC types of
potential concern, including: the basis of these EC
Naturally produced compounds such as toxins produced by fungi,
bacteria and plants;
Bio-terrorism/sabotage agents;
Human personal care products such as essential oils, herbal
medicines, antibacterials and fragrances;
Agricultural Contaminants
Emerging persistent organic pollutants such as flame retardants and
dioxin-like compounds;
Veterinary medicines such as antibiotics and antiparasitic agents;
Hormones such as synthetic and natural estrogens and androgens;
Nanomaterials;
Human medicines;
Metabolites and transformation products of man-made chemicals
that are produced from biological, chemical and physical breakdown
reactions
Sources of Agricultural Contaminants

• Emerging contaminants will be released to the agricultural

environment via a number of routes:

Veterinary medicines and their metabolites will be released directly

to soils (animals at pasture) or indirectly when manure and slurry
from intensive livestock facilities is applied to agricultural land as a
fertiliser
Sources of Agricultural Contaminants

Human pharmaceuticals and personal care products will be released

through the application of sewage sludge (biosolids) to land or from
irrigation with wastewater effluent

Other ECs may be formed in the environment itself

Sources of Agricultural Contaminants Figure 2. Routes of entry of ECs to the agricultural environment

Human use Livestock use Plant protection

Pharmaceuticals Hormones Veterinary Nanomaterials
Personal care products medicines
Nanomaterials Metabolites
Flame retardants
Metabolites

Excretion to Application
soils
Waste or
Wastewater Manure
treatment storage

Sludge/biosolid
Application ; wastewater Manure
Irrigation; compost application
application etc.

Formation Formation of
of transformation Agricultural soils natural toxins
products
Techniques – Emerging Pollutants
• Techniques for identifying and quantitating emerging contaminants:

Gas chromatography-mass spectrometry (GC-MS),

LC-MS, or tandem mass spectrometry (gas chromatography-mass
spectrometry/mass spectrometry (GC-MS/MS),
LC-MS/MS and gas chromatography x gas chromatography-mass
spectrometry (GCxGC-MS) methods are at the forefront
Techniques – Emerging Pollutants
• N.B.: Liquid chromatography & gas chromatography techniques,
coupled with mass spectrometry, provide extremely powerful
analytical tools by combining the intrinsic properties of the individual
techniques

• As such, the major research has been on the improvement of various

facets of these analytical techniques for the eventual development of
robust environmental monitoring systems
Major Challenges

• N.B.: One of the major challenges in the environmental analysis of

emerging contaminants is that, due to the number of parent
contaminants, a great number of metabolites, degradation &
transformation products of unknown toxicity & persistence is
expected to exist
Traditional Microbiological Testing Techniques

• The traditional strategies for routine microbiological testing include

gram staining, colony morphology, microscopic examination,
differential growth on selective media & various biochemical tests
(catalase and oxidase tests), with either manual or automated
methods or – in some cases – commercial kits
Traditional Microbiological Testing Techniques

• Furthermore, secondary phenotypic characterisations complete the

microbial identification process

• There are several drawbacks to conventional culture assays as routine

& robust detection tools for pathogens
Traditional Microbiological Testing Techniques

• The major drawback is that the analyses are slow (they can take
between two and seven days) & labour intensive as pathogens need
to be cultured & enriched in selective media to isolate specific
pathogens from other microorganisms
Traditional Microbiological Testing Techniques
• Moreover, in many instances, pathogenic concentrations may be too
low for cultural detection, but may still be high enough to cause
infection

• Some studies have even highlighted the fact that the microbial load in
water can be significantly underestimated using the traditional plate
count method due to the presence of physiologically active bacteria
that are unable to form colonies on culture media
Polymerase Chain Reaction (PCR)
• Molecular diagnostics are better alternative approaches to culturing
techniques for identifying pathogens

• PCR enables rapid bacterial identification by targeting conserved

genes such as those coding for the ribosomal RNA (rRNA) of
pathogens

• The PCR techniques have several advantages over culturing methods

Advantages of PCR
• Firstly, PCR allows for the identification of slow-growing organisms
and has been used to establish pathogenesis for uncultivable
organisms

• The PCR has been used successfully to detect the presence of H.

pylori DNA in drinking water

• Secondly, results are generally obtained within a short time, especially

if real-time PCR (qPCR) is used
Advantages of PCR
• Real-time reverse transcriptase PCR (qRT-PCR) uses specific probes
that generate significant information on the presence, quantity &
distribution of classic & new emergent pathogens in water with a high
level of sensitivity & specificity

• The qPCR displays better specificity, sensitivity & reduced time

requirements compared with available culture-dependent methods &
has been widely & routinely used to directly detect pathogens in
research & clinical diagnosis
Disadvantages of PCR
• One disadvantage of these molecular techniques is the targeted
approach for specific microbial genera or species…therefore, multiple
pathogens can’t be monitored at a time

• Even though PCR is a very sensitive detection technique, it faces

challenges with viral identification…due to low concentration of viral
particles in environmental water & their extraction procedures
Pyrosequencing techniques
• Pyrosequencing technology is a revolutionary technique based on
DNA sequencing, utilising enzyme-coupled reactions &
bioluminescence to monitor the pyrophosphate release that
accompanies nucleotide incorporation

• Unlike PCR, where scientists are limited by known sequence

information and must select the pathogens to be considered in each
assay, a high-throughput sequencing approach is unbiased and makes
it possible to detect novel pathogens
Pyrosequencing techniques

• Sequencing technology also has the potential to provide an unbiased

detection approach for waterborne pathogens with a single common
protocol
Mass spectrometry techniques
• Mass spectrometry has emerged as a powerful tool for analysis and
proteomics research and the first attempts at utilising it for the
characterisation of organisms were made in 1975

• The soft ionisation Matrix-assisted Laser Desorption Ionisation time

of flight mass spectrometry (MALDI-ToF-MS) was particularly useful
for large biomarkers

• As such, MALDI-ToF-MS can be used to characterise a wide variety of

microorganisms, including bacteria, fungi and viruses from water
Mass spectrometry techniques

• MALDI-ToF-MS identifies microorganisms by analysing the total

protein and generating mass spectra from whole cells and their
comparison to reference spectra

• Two general approaches are used when characterising

microorganisms using MALDI-ToF-MS
Mass spectrometry techniques
• The first approach is fingerprinting intact microorganisms where
intact cells are used to generate unique spectral fingerprints that can
be compared with previously collected fingerprints

• This is because spectral fingerprints vary between microorganisms

and the spectra obtained are reproducible if the bacteria are grown
under the same conditions

• This approach is relatively simple, as it is possible to use minimally

processed intact cells
Mass spectrometry techniques

• The second approach is the bioinformatics-enabled approach often

referred to as MALDI-ToF-MS biotyping (MTB)

• Here, masses associated with an unknown microorganism can be

identified by comparing them with masses of proteins in protein
databases