Lake Geneva Monitoring Program

2203ENV Environmental Chemistry and Monitoring
Lake Geneva Monitoring Program
Regina Kimble: s5000977

Tyler Hofmann: s2967975
Tish King: s2839375
Charles Pinson: s2937832
Contents
Setting Monitoring Program Ejectives
Define the Issue
Define Information Requirements
Compile Available Information
Set Objectives
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3
3
6
Study Design
Determine the Study Scope
Consider Sampling Design Issues
Specific Data Requirements
Budget
6
6
7
8
Feld Sampling Program

Sample Collection Methods
Sample Container Requirements for Analyses
Sample preservation and Storage Requirements
Field Measurements Needed
Quality Control
Occupational Health and Safety
9
10
10
10
11
11
Laboratory Analyse
Identify Desired Analyses
Select Appropriate Analytical Methods for Required Detection
Limits and Precision
Undertake Analyses with Appropriate Quality Assurance and Quality Control
Occupational Health and Safety
12
12
13
14
Data Analyses and Interpretation

Data Preparation
Data Integrity
Data Analysis
Relation to Study Objectives and Conceptual Model
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15
15
16
Reporting Information Dissemination
16
Reference List
16
Setting Monitoring Program Objectives

Define The Issue
Lake Geneva is one of the biggest lakes in Western Europe, with a length of 72.5km, and
crosses the borders of Switzerland and France (CGN, 2013). Lake Geneva is surrounded by
cities Geneva and Lausanne, which are some of the most populous cities in Switzerland
(CIPEL, 2014). Water from the lake is used as drinking water for over 800,000 of Genevas
residents (CIPEL, 2014). The river is also utilised for swimming, other water recreational
activities and fishing (CIPEL, 2014). Its essential that the water in Lake Geneva is monitored
for harmful substances.
Lake Geneva receives most of its water from urbanised river sources (Faure et al., 2015)
and over 60% of its banks are heavily affected by human interactions (CIPEL, 2014)
Accumulation of microplastics in the lake, largely caused by runoff from densely populated
cities, is a major threat to many species which inhabit it (Faure et at., 2015). Microplastics
(<5 mm) have the ability to adsorb harmful chemical compounds that can bioaccumulates up
the food chain (Reisser et al., 2013). Persistent organic chemicals have hydrophobic
qualities which have shown to adsorb onto plastics in the ocean (Seltenrich, 2015). Plastic
particles from the ocean have shown to possess a higher concentration of pollutants
compared to the water where it was found (Seltenrich, 2015). Since microplastics are so
small, marine animals ingest them without even trying (Seltenrich, 2015). This can
bioaccumulate and potentially affect humans (Reisser et al., 2013).
Define Information Requirements
The stakeholders for this monitoring program are the Switzerland federal government,
French government, product manufacturers of microplastics, environmental groups, local
fishing industries and ecotourism. This is due to the duel-national and local dependency on
the lakes fishing and recreational resources (CIPEL, 2014).
Compile Available Information
There are major cities that lie near Lake Geneva: Geneva, Lausanne, Montreux and
Thonon-les-Bains. Urbanised rivers feed into Lake Geneva: Rhone, Aubonne, Venoge,
Dranse and Vuachere River (CGN, 2013). The Rhone drains 18% of the waters and
provides most of the lake water for Switzerland (CGN, 2013). From France, the Dranse is
the second biggest provider of water to Lake Geneva (CGN, 2013). It also receives sewage
discharge from a Lausanne treatment plant (Monna et al., 1999). More than 150 professional
fisherman use Lake Geneva, catching perch, whitefish, trout, char and pike (CIPEL, 2014).
Lake Geneva is separated into three different parts (Figure 1), due to different types of
formations: the Petit-lac, the smallest part from Nyon to Geneva; the Grand Lac, the biggest
and deepest part from Evian and Lausanne and the Haut-Iac, the upper eastern part of the
lake, defined by Vevey-Montreux-Bouveret-Thonon (CGN, 2013).
Figure 1: Picture of Lake Geneva and how it's separated into three different parts.
(CIPEL, 2014)
The six main types of microplastics found are polypropylene, polyethylene, polyvinyl
chloride, polyethylene terephthalate, polyurethane and polystyrene (PlasticsEurope, 2013).
Small microplastics (<1 mm) adsorb harmful pollutants such as 12 PCB congeners, 16
polycyclic aromatic hydrocarbons, 19 organochlorine pesticides (Faure et al., 2015).
Additives like phthalates, bisphenol A, polybrominated diphenyl ethers are also adsorbed
and have been found in marine animals, like the myctophid fish in Tasmania (Reisser et al.,
2013). They absorb chemicals more efficiently than larger microplastics because they have a
larger total surface area, but its not fully understood what other characteristics make them
more susceptible to adsorption (Faure et al., 2015). Studies showed that bird pellets exhibit
short and long-term consumption of chemicals adsorbed from microplastics and fish
dissections showed signs of microplastic ingestion (Faure et al., 2015). The amount of small
microplastics in the surface water of Lake Geneva was 5.6 times bigger than larger
microplastics (Faure et al., 2015). It was also found that more rainfall, downstream sampling
and gyers lead to higher concentrations of microplastics (Faure et al., 2015). The effect of
vertical mixing and trends in microplastic accumulation havent been fully studied. There isnt
a standardized method of sampling and classifying microplastics, or a precise way of
analysing the chemicals adsorbed.
During the 1960s through the 1980s, Lake Geneva experienced a high pollution input from
sewage treatments that lead to eutrophication, which almost killed its fish population (Monna
et al., 1999). The residence time of water in Lake Geneva is 11.3 years and today is
considered clean enough to do recreational water activities in (CIPEL, 2014).
Recent studies have been done on the abundance of microplastics in Lake Geneva, and its
abundance have been found to be higher than expected, however further studies need to be
done to assess the impact this has on marine species and how it ultimately affects humans
through the food web (Faure et al., 2012).
Monitoring is currently done on physico-chemical properties, biological properties and on

micropollutants, which are pollutants that have the ability to bioaccumulate (CIPEL, 2014).
While micropollutant concentrations are being monitored, its effects on marine species and
origins are not being investigated. This is important because having quantitative data on how
micropollutants enter the lake and are magnified in marine species can help mitigate its
potentially harmful effect on humans.
Table 1 measurements were collected after rainfall occurrences, which have been known to
increase the abundance of microplastics (Faure et al., 2015). Table 2 shows how urbanised
rivers contribute to microplastic densities in Lake Geneva.
Table 1: Densities of macro and microplastics in surface waters of Swiss Lakes.

(Faure et al., 2015)
Conceptual Process Model and System Understanding
Figure 2: Conceptual process model for Lake Geneva.

(UNEP Year Book, Kershaw et al. 2011)
Microplastics are inputted into lakes through urbanised river sources (Faure et al., 2015) .
Lake Geneva has 5 main river sources that are linked to populated cities (Faure et al.,
2015). In Figure 2, sources of microplastics come from wind and water litter (gray arrows).
Microplastics are transported through the marine environment by the water column and
through processes like sedimentation (orange arrows). Microplastics are taken up by the
marine environment through animals and placement in the sediment (black arrows).
Disposable plastic packaging abrasion and microbeads from body products are major
contributors to microplastics which are sourced from urban areas (PlasticsEurope, 2013).
Rainfall (not depicted in figure 2) is one of the major environmental events that can
contribute to the abundance of microplastics in lakes.
An increase in microplastics would cause many undesirable effects for Lake Geneva and its
marine animals. These include an increase in microplastics in filtered water, as they are too
small to be effectively taken out during treatment, and bioaccumulation of harmful pollutants
in fish (Seltenrich, 2015). Both of these effects directly impact human health. Without
monitoring microplastics, an increase in its density would cause fish to be toxic to humans
and affect 800,000 people in Geneva who depend on Lake Geneva for drinking water.
Set Objectives
The objective of this monitoring program is to assess the concentration of microplastics in
surface waters of Lake Geneva due to the importance of Lake Genevas safety as its an
important water body in Switzerland and France. The effects of adsorbed pollutants in fish
will be assessed to see the potential effect on human consumption. The correlation between
microplastic density and concentration of pollutants in fish will be investigated. The results
will be given to the stakeholders to take action if need fit.
Study Design
Determine the Study Scope
The investigative monitoring program will carry on for six months and take place in Lake
Geneva and its surrounding major river inlets. The samples will be collected as per the
sampling design throughout the lake and its corresponding rivers.
Consider Sampling Design Issues
Field Sampling Sites & Spatial Variability
A systematic judgemental sampling technique is used to consider the spread of
microplastics along surface waters of Lake Geneva. Five trawls for three kilometers will be
conducted for major inlets that run through Lake Geneva using a floating manta trawl net
with a 90 x 30-cm opening and a 0.333-mm mesh. To avoid mixing in the surface trawls, the
trawl net will be on the downwind side of the boat. Judgmentally, a trawl near the inlet rivers
will be conducted running for three kilometers. This will show urban influences on the
concentration of microplastics and how microplastics are distributed within the lake. Fish will
be randomly collected in where the trawls took place. Sixty fish of five different species
(perch, whitefish, trout, char and pike) will be caught.
Frequency
Since its known that weather conditions have an effect on the concentrations of
microplastics, two different set of trawls in each allocated area will be conducted. One after a
heavy rainfall, one on a mild weather day and one on a windy day. Sampling on a mild day
will indicate the normal concentrations of microplastics. Sampling after a heavy rainfall will
show how it increases the concentrations of microplastics. As wind is a variable that hasnt
been fully investigated, the windy day trawl will add data and knowledge to that. In total,
there will be 30 samples taken over the six months. Fish will be caught in April and
November. 6 fish, of the species mentioned previously, will be caught at each location, for
each month. In total, 60 fish will be caught. Fish spawn in March, so juvenile fish are
included in the study. A six month gap will allow for growth and more variation in fish
characteristics.
Precision and Accuracy
During sampling, its important to make sure methods and collection are precise and
accurate, as this ensures the data is of high quality and defensible in legal cases. Design
issues concerning precision and accuracy are highly correlated to the parameters discussed
above. This correlation occurs because precision and accuracy is dependent on frequency
of samples per location and sampling methods. The samples in this monitoring program will
be compared to other surface trawls done in Lake Geneva, to show accuracy. Doing trawls a
second time in each location will take precision into account. By considering both accuracy
and precision in the sampling methods, it ensures the data collected will be of a high enough
quality to use in an analysis report.
Specific Data Requirements
The monitoring program in Lake Geneva will measure the concentration of microplastics,
concentration of adsorbed pollutants within the microplastics and the affects they have on
native fish species.
Concentration of Microplastics
Microplastics are smaller than five millimeters and will be put into different classes based on
appearance (colour, shape, density, etc.) and possible origin. The chemical composition of
microplastics will be determined using infrared attenuated total reflectance analysis.
Microplastics bigger than five millimeters will be catalogued as they are a source for smaller
microplastics.
Adsorbed Pollutants in Microplastics
Smaller microplastics adsorb harmful chemicals that bioaccumulate in the food chain and will
be identified by gas chromatography-tandem mass spectrometry. 2 PCB congeners, 16
polycyclic aromatic hydrocarbons, 19 organochlorine pesticides and additives phthalates,
bisphenol A, polybrominated diphenyl ethers will identified in the gas chromatographytandem mass spectrometry test .
Adsorbed Pollutants and Microplastics in Marine Animals
Microplastics are so small and this causes marine animals to unconsciously ingest them
(Seltenrich, 2015). Larger fragments will be identified during dissection and the adsorbed
pollutants mentioned above will be tested using gas chromatography-tandem mass Field
Saspectrometry with tissue samples from the gut of the fish samples.
Budget
The costs to monitor Lake Geneva have to be investigated. As mentioned before, a
judgemental systematic sampling will be used to collect surface microplastics every 3
kilometers for 5 trawls, under 3 different weather types, with each trawl repeated 2 times.
The costs for the field expenses will be covered by Griffith University.
The sorting of microplastics will be done with equipment from Griffith University and will
undergo infrared attenuated total reflectance analysis to infer the chemical composition. The
cost of this is based on prices from University of Michigan and is outlined in table 2.
Analytical Test
Cost per Hour
Number of Hours
Total
Microplastics
$86.76
48
$4164.44
Table 2: Costs for infrared attenuated total reflectance analysis to assess chemical
composition of microplastics. All costs are in AUD.
The pollutants adsorbed onto these microplastics, mentioned previously, will be tested using
gas chromatography-tandem mass spectrometry. 15 samples will be tested. Each trawl is
conducted 2 times, the sample of those for each weather condition will be combined for
every location.
60 fish of five different species (perch, whitefish, trout, char and pike) will be caught and
gutted out to see the amount of microplastics, using equipment from Griffith University.
Tissues from the gut will be analysed through gas chromatography-tandem mass
spectrometry for the same pollutants. Table 3 outlines the costs for gas chromatographytandem mass spectrometry based on quotes from Montana State University.
Analytical Tests
Cost per sample
Number of samples
Total
Fish gut tissues
$66.66
60
$3,999.60
Lake microplastics
$66.66
15
$999.90
Total
$133.32
75
$4,999.50
Table 3: Costs for gas chromatography-tandem mass spectrometry analysis for microplastic
and fish tissue samples. All prices are in AUD.
In total, the cost of this monitoring program on Lake Geneva for a period of 6 months adds
up to $9,163.94. Transport, containers and miscellaneous equipment like gloves and paper
towels will be covered by Griffith University.
Feld Sampling Program

Sampling Collection Methods
Since lake Geneva is one of the biggest lakes in western Europe, a systematic judgement
sampling technique will be implemented in order to achieve an accurate representation of
the whole system. This will help account for the spread of microplastics along the surface of
the lake. Five trawls over three kilometers will be conducted for each major inlet and outlet
that runs through Lake Geneva by using a floating manta trawl net (shown in figure 3). This
is the conventional method for collecting fine particles in water. The net has a 90 x 30 cm
opening and 0.333 mm mesh. To avoid mixing in the surface trawls, the trawl net will be on
the downwind side of the boat. Fish will be caught in a conventional fishing net while the
trawls take place. A total of sixty fish will be collected. This will be made up of six fish from
each of the five locations and the test will be conducted once in April and once in November
where the trawl will be caught for analysis.
Since weather conditions are known to have an effect on the concentration of microplastics,
three different set of trawls in each of the five allocated areas will be conducted. One trawl
will be conducted after a heavy rainfall, one on a mild weathered day, and one on a windy
day. Sampling on a mild day will act as a control in accordance to the Australian and New
Zealand Guidelines for Fresh and Marine Water Quality (ANZECC, 2000). This will indicate
the normal concentrations of microplastics. Sampling after a heavy rainfall will show how it
increases the concentrations of microplastics. As wind is a variable that has not been fully
investigated, the windy day trawl will serve to gather incomplete data and knowledge. There
will be a total of 30 different trawls. Fish will be caught in April and November. Fish spawn in
March, so juvenile fish would be included in the study. A six-month gap will allow for growth
and more variation in fish characteristics.
Figure 3: The conventional in-water setup for a manta trawl. Image from (NOAA Marine
Debris Program, 2016)
Sampling Container Requirements for Analytes

In accordance with the Industrial waste resource guidelines: Sampling and Analysis of
Waters, Soils, and Waste, the sample containers should have minimum interaction with the
samples. For this reason glass sample containers will be used to store the plastic collected.
This will ensure that the container will not chemically react to the analyte, nor will the small
microplastics stick to the insides of the container as they would if it were made from plastic
(EPA Victoria, 2009).
A 100L esky filled with with a 1:1 ratio of water and ice will be used to transport and store the
fish. For each day that the fish are kept on ice, some of the water should be drained and the
ice replenished to keep the fish fresh (Blessing, Marshall and Balcombe, 2010).
Sample Preservation and Storage Requirements
Storage requirements differ between the microplastic samples and the fish samples. After
completing each trawl, carefully rinse the outside of the manta net with neutral water before
emptying it. This will remove particles that could bias the sample (NOAA Marine Debris
Program, 2016). Empty the contents onto a 0.333 mm mesh sieve and rinse the basket
again to ensure all that no particles are stuck inside. Before removing organic litter such as
leaves and sticks, thoroughly rinse them to collect any small attached particles. Once
complete, the particles from the sieve are placed into glass sample containers which will
then be labelled with the location and date of the trawl. Once this is complete, keep samples
containers at room temperature to help protect the integrity of the analyte (NOAA Marine
Debris Program, 2016).
Sample preservation of the fish involves placing the freshly caught samples into a large esky
filled with an ice-slurry. A study conducted by Griffith University in Nathan titled Humane
killing of fish for scientific research: a comparison of methods (Blessing, Marshall and
Balcombe, 2010) found that this was the most humane way to euthanise fish. The low
temperature slows the heart rate putting them into a state of unconsciousness and shortly
after this the heart will stop beating (Blessing, Marshall and Balcombe, 2010). The fish will
need to be kept in this slurry until they are analysed, in order to slow the decaying process.
However, it will not completely freeze the fish and for this reason the samples must be kept
out of the sun. They must be taken to the lab for examination immediately so the fish will not
start to rot and the ice must be replenished regularly (Blessing, Marshall and Balcombe,
2010).
Field Measurements Needed
In regards to the nature of the experiment, most observations are made in the laboratory.
However, it is necessary to record how full the lake is (the percent of capacity) and make
accurate field notes, such as: date, time, other observations such as wind direction, visual
observations such as rubbish, possible sources of the pollutants, sample coordinates and
photos of the sampling process (ANZECC, 2000).
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Quality Control
Effective quality control management will ensure efficiency and accuracy for the project.
OH&S procedures and data management plans must be in place before sampling begins. A
field book will be used throughout the sampling process to ensure that all relevant in situ
information is recorded. The weather must be recorded in the week leading up to the trawls
because some trawls are to be conducted after a moderate rainfall event (NOAA Marine
Debris Program, 2016).
A Beeline GPS system will be used to ensure the correct positioning of and length of trawl
transects. The coordinates of the sample site will be recorded in the field book so that the
exact location can be found for the future sample events (NOAA Marine Debris Program,
2016). The naming the samples should be standardised between the datasheets and the
sample labels. All of the procedures from trawling to data processing must be monitored for
consistency (ANZECC, 2000).
While trawling, ensure that the manta net is being towed correctly. The net should not be
splashing around while in tow (this would impact the sample integrity) but it should be
skimming the water smoothly along the surface. If the net is not towing correctly, the speed
of the vessel may need to be adjusted to provide optimum net positioning in the water which
allows the maximum amount of water to pass through the net (NOAA Marine Debris
Program, 2016). Special care is to be taken when rinsing debris by using deionized or
filtered water in a controlled environment (ANZECC, 2000). This will help ensure accurate
results. If possible, a second person supervising the process can help remove human error
(ANZECC, 2000). Qualified personnel must calibrate all instruments. Cleaning and
inspections must be undertaken before and after each sample process to help ensure
consistency and precision (ANZECC, 2000). The three different trawls of the same sight will
be compared to asses if there is any variation caused by weather effects.
Occupational health and safety
For this study, safety is the main priority because the task involves working outdoors where
there are a number of safety factors that are inherited and can not be controlled. However,
with proper management the risk level can be kept to a minimum. To manage OH&S, a risk
assessment documenting all potential hazards during the sample process will be complete.
The potential hazards will be encountered: while loading equipment into vehicle, driving to
the sample sight, being on the boat, taking the samples, equipment handling, hazardous
exposure (weather, UV rays, and dangerous substances), and encounters with flora and
fauna (ANZECC, 2000). The risk assessment will include: how to travel to the site safely
(rest every two hours, dont drive under the influence of drugs and alcohol, ect), correct lifting
procedures, ensuring personal protective equipment is available and worn (closed in shoes,
long sleeve shirt, life jacket), ensuring a first aid kit is present, and that there is always more
than one person present when doing the sampling or travelling. All staff involved in the
sampling process must be qualified to do so, must complete the mandatory safety inductions
and have first aid qualifications to ensure that they are aware of potential hazards and how
to manage them (ANZECC, 2000).
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Laboratory Analysis
Identify Desired Analytes
The laboratory analysis will take place in two parts. The first will determine the total amount
of microplastics present in the surface waters that were sampled in Lake Geneva. Part two
involves dissecting the fish to reveal the amounts of microplastics that have been ingested
by them. For both parts, the analytes are the microplastics themselves, which will be
identified and separated into their respective groups.
Select Appropriate Analytical Methods for Required Detections Limits and Precision
Surface water samples:
The 30 samples were stored at 4oC in 100mL sealed glass jars until analysis. Glass was
used through the entire analyses of both the waters and the fish to avoid contamination by
abrasion of plastic containers). The samples were centrifuged to separate the plastics from
the water. The sample was then dried in an oven at 50oC for 2 hours, having been taken out
and shaken periodically. The dried microplastics were then ran through a 300m sieve and
separated into three different sizes: >5mm (macroplastics), >1mm (large microplastics) and
>300m (small microplastics). The macroplastics were identified (distinguished from organic
matter and cell matter) under a stereomicroscope and catalogued. Both the large and small
microplastics were further dried at 500C and underwent wet peroxide oxidation to dissolve
organic matter (OM) with a 35% H2O2 with 0.05M FeII catalyst for 6 hours (Adapted from
Faure et al., 2015). Once dried, the mass of microplastics was recorded. The mass of
organic matter (including plankton and cell material) was recorded as a reference used to
contrast the proportions of plastics and OM. Knowing the volume of water sampled by the
trawls, the results could be extrapolated to both particles and mass per litre (number L-1 and
mgL-1) and conclusions could be drawn as to how meteorological factors affect the presence
and abundance of microplastics.
Fish dissections:
The 60 fish were stored on ice until analysis. Once thawed out, their guts were removed and
the contents rinsed and ran firstly through a 5mm sieve, repeated three times to ensure
everything <5mm passed through and then a 300m sieve. From here on, the same
methods were used as the plastics from the water samples.
Plastics from each fish species and location were kept in separate jars until they could be
analysed.
Identification of plastics:
For each size (>5mm, >1mm and >300um) the plastics were separated according to their
characteristics (colour, shape, density etc.) and possible origin (cosmetics, fishing lines,
textiles, polystyrene packaging etc.). Out of the thousands of pieces of plastic that were
collected, 100 of each size and type were picked at random to undergo infrared attenuated
total reflectance (IR ATR) spectroscopic analysis to identify which of the 6 possible plastic
polymers they were according to their spectral peaks (Adapted from Faure et al., 2015).
Then 10 microplastics from each fish will undergo IR ATR to identify their chemical
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composition. Despite being costly and time consuming, IR ATR is still the most effective
method of identifying the chemical composition plastics.
Once identified and catalogued, the plastics were analysed using gas chromatography
tandem mass spectrometry (GC-MS) to identify which, if any, harmful compounds or
additives which had been absorbed into them (Methods adapted from Hirai et al., 2011). Gas
chromatography allows you to find out how many chemical compounds are in the sample,
while mass spectrometry determines what the chemical formula for each compound is (Hirai
et al., 2011). The compounds that were tested for were PAHs, PCB congeners,
organichloride pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs) as well as
the additives phthalates, nonylphenols and bisphenyl A. The following blanks for the GC-Ms
and LC-MS were required:
3x 500mg LDPE bands cleaned with DCM and methanol
3x method blanks with extraction into an empty tube,
4x instrument blanks of 10mL DCM reduced to 1mL.
The limits of detection (LOD) and quantification (LOQ) were calculated as +3 and +10
respectively.
Lab Analysis
Undertake analyses with appropriate Quality Assurance & Quality Control
A comprehensive fieldwork Quality Assurance (QA) and Quality Control (QC) plan will be
implemented. This is to evaluate the skills at the end of training and during data collection.
Included will be applying validation (call-back) effort to detect falsification (Queensland
Government, 2016). Field blanks, trip blanks and laboratory blanks are important in the
monitoring program when taking samples in the field and analysing the data in the
laboratories (Queensland Government, 2016). The field blanks will have a sample of analyte
free water poured into the container in the field, preserved and transported appropriately to
the laboratory with field samples. This will assess contamination from field condition during
sampling. Trip blanks will be clean samples of a matrix taken from the laboratory to the
sampling site. They will be analysed only for volatile compounds and then transported back
to the laboratory without having been exposed to sampling procedures. They will be
transported in a cooling container. This will assess the contamination that may be introduced
during shipping and field handling procedures.
The laboratory blanks that will be used will be method blanks and instrument blanks. Method
blanks will be prepared to represent the matrix as closely as possible. It will prepare, extract,
digest and analyse exactly the same way as the field blanks (Queensland Government,
2016). This will consider whether contamination has been introduced during sampling
preparation. Instrument blanks will be analysed with the field samples. It will assess the
presence of instrument contamination, but additionally whether instrumentation is absent.
The amount of blanks will be defined by the analytical method or the analysts discretion,
especially after concentration samples.
Various samples from each field site will be collected and analysed. Each replicate sample
will be used to establish a different weather condition. Duplicates will establish how well
each sample was performed at each site and determine errors, such as contamination.
Information that is relevant to field and laboratory sampling will be recorded in detailed log
books (Queensland Government, 2016). Each sample taken from each site will be recorded
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with detailed information, and will include observations on the surrounding areas, condition
of the lake and weather conditions. Inclusively, videos and photos will be documented as
well as maps of each site. Coordinates of each site will be recorded and will aid in collecting
samples from the same site. This will decrease error in the method of the collection of
samples. All glass bottles will be labelled accordingly to the site, and replicate with date and
time with the person who collected the samples. Additionally, a record of everyone who
handles the samples will also be documented throughout the entire sampling, transport and
analysis (Queensland Government, 2016).
Consideration of contamination and validation is important. To maintain the integrity of the
samples, they will be transported to the appropriate laboratories via appropriate
transportation methods. Sample jars will be packaged in sample carrier boxes to minimise
risk of breakage, leakage or spillage. Security measures will be put in place and
documentation will accompany the samples. All samples will be analysed within the
maximum holding times specified. It will be essential to re-recheck caps on the glass bottles
to make sure they are tight and properly sealed. Glass bottles will also be separated from
each other to prevent physical contact to elude breakage. All samples will be labelled with
FRAGILE, HANDLE WITH CARE or THIS SIDE UP. This will minimise movement
(Queensland Government, 2016).
Occupational health and safety

Acknowledging Occupational Health and Safety is essential when working in the
laboratories, thus it is recommended to identify all possible hazards involved in data
collecting and data analysis with a specific safety plan being developed for each monitoring
plan. All potential hazards involved in the laboratory analysis will be identified and
documented with a risk assessment and hazard control measures. Common hazards in the
laboratory will include: animal, biological, decontamination chemical, analytes or stabilisation
for acidification and physical equipment handling. When conducting a sampling event, the
right safety equipment will make the task safer. This equipment can be preventative or
provide assistance in the case of an incident. A checklist will be put in place to make sure
these risks are avoided.
Preparation of laboratory safety training will be applied, with all procedure and associated
safety information displayed in the appropriate areas. Accident or emergency situations
involving hazards will have written instructions for a response procedures and protocols.
Appropriate behaviour will only be tolerated, thus conducting responsible and professional
matter is an obligation. Personal Protective Equipment (PPE) is important when in the
laboratory. Laboratory coat and appropriate footwear will avoid skin exposure and protect
risks of potential splashes. Safety glasses worn at all times and latex gloves when handling
biological and chemical apparatus. Hair must be tied back and jewellery not worn to elude
anything that may catch on any equipment. No food, drink beverages and chewing gum to
be present in the laboratories. If using chemicals, knowledge of the chemical fume hood or
biosafety cabinet must be understood, if not training will be provided. Maintaining good
house cleaning (aisles clear) will reduce risks of any potential hazards. Active experiments
are not to be left unattended, especially if being heated, or visibly reacting. Importantly, good
personal hygiene before entering and exiting the laboratories will decrease contamination.
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Data Analysis and Interpretation

Data Preparation
To make sure that the samples and data collected from Lake Geneva can be analysed
correctly, every sample will have a specific code to identify it, along with information on
where it was taken from and what weather condition it was taken in. The data collected will
be out into glass jars to avoid contamination. This data will undergo statistical analysis like
scatter plots, histograms, box plots, standard error plots and Q-Q plots to show patterns and
trends in the data. Box and whisker plots will be graphed to compare the median abundance
of microplastics for each location with each weather pattern to see if theyre normally
distributed or have any outliers. The concentrations of microplastics will be plotted against
the concentration of adsorbed pollutants in fish, at respective locations, to see if theres a
correlation between the two. The graphical representation mentioned above will be done on
raw data and data that has gone through statistical tests. The graphical representations will
also show if the data meets assumptions needed to do statistical tests on it (SPSS, 5th
edition). If it does not meet those assumptions, the data can be manipulated.
Data Integrity
To make sure the data is sound, the methods used will be checked by comparing it to
previous studies done, general procedures of Quality Assurance, known concepts and
principles. This incorporates the Quality Assurance (QA) and Quality Control (QC)
mentioned previously to ensure our data is precise and accurate (Queensland Government,
2016). Factors to be stated in a policy statement will include a manual of QA and QC,
standard operating procedures of in the field and in the lab and definitions of QA and QC.
Other definitions of quality control terms will also include: test portion, replicate, duplicates,
spiked samples, laboratory control samples, control charts and statistical process control
(Swiss, 1992). Calibration will be discussed in regards to the analysis including method
detection limits, limits of quantitation and linearity. The data will be checked for outliers and
missing data points, which could be a result of human error.
Data Analysis
The data collected from this monitoring program can be used to determine what sources of
rivers input the most microplastics into Lake Geneva and how this affects bioaccumulation of
adsorbed pollutants and microplastic abundance in fish. To determine this, statistical tests
like t-test and ANCOVA f-tests will be used. The measurement parameters include
abundance of microplastics, types of microplastics and concentration of pollutants adsorbed.
The concentration of pollutants adsorbed in the fish do not necessarily mean they came from
the microplastics, but this will be investigated using scatterplots mentioned previously and
statistical tests like linear regression to see if the two variables are dependent on each other.
The windy day data will also be checked to see if it has a significant difference from mild
weather and rainfall events, as windy day data on microplastics has not been previously
investigated. The significance of weather conditions on the concentration of microplastics will
be investigated using multiple regression to see if adding or taking away the weather
conditions has an overall effect on the concentration of microplastics at each location.
15
Relation to Study Objectives and Conceptual Model

After the data undergoes statistical tests, the data will be checked to see if the main
objectives were reached. The results from the analysis will determine if different river
sources and weather patterns affect the concentration of microplastics in Lake Geneva.
Also, the results will determine if an increase in microplastics means an increase in
adsorbed pollutants in fish. The data will also be compared to the conceptual model
mentioned previously. If the data does not support the objectives or the model, the
monitoring programs direction may need to be revised or the monitoring program may need
to be conducted again.
Reporting and Information Dissemination
The information collected from this monitoring program will be put into reports for the
stakeholders mentioned previously. A non-theoretical report that focuses on the maths and
methods used will be sent first, to see if it complies with the stakeholders requirements and
needs. It will also be available so other scientists can review the methods and maths done
on the data to check its correctness. After this, the data will be incorporated with the theory
on microplastics and how they bioaccumulate, and this report will be sent out to the scientific
community and the stakeholders. The stakeholders can use these reports to determine if
Switzerland and France should ban microplastics like other countries. The information from
this monitoring program could be used to regulate microplastics and provide a model for
other countries to monitor their lakes for microplastics.
References
Australian Government, Department of Environment, 2000. Australian and New Zealand
Guidelines for Fresh and Marine Water Quality. Volume 1. [online] Canberra: Australian and
New Zealand Environment and Conservation Council, pp.Chapter 1-7. Available at:
<https://www.environment.gov.au/system/files/resources/53cda9ea-7ec2-49d4-af29d1dde09e96ef/files/nwqms-guidelines-4-vol1.pdf> [Accessed 5 May 2016].
Blessing, J., Marshall, J. and Balcombe, S., 2010. Humane killing of fishes for scientific
research: a comparison of two methods. Journal of Fish Biology, 76(10), pp.2571-2577.
CGN, (2013). Lake Geneva. [online] Cgn.ch. Available at: http://www.cgn.ch/engb/content/cgn/l%C3%A9man.aspx [Accessed 5 May 2016].
CIPEL, (2014). FICHE SIGNALTIQUE DU LMAN ET DE SON BASSIN VERSANT.
[online] CIPEL. Available at: http://www.cipel.org/wpcontent/uploads/2016/02/Fiche_signaletique_Leman_2015.pdf [Accessed 3 May 2016].
Commission internationale pour la protection des eaux du Lman, (2014). Caractristiques
du Lman. [online] Cipel.org. Available at: http://www.cipel.org/le-leman/caracteristiques/
[Accessed 5 May 2016].
16
Environmental Protection Authority Victoria, 2009. SAMPLING AND ANALYSIS OF

WATERS, WASTEWATERS, SOILS AND WASTES. [online] IWRG701, pp.3-4. Available at:
<http://www.epa.vic.gov.au/~/media/Publications/IWRG701.pdf> [Accessed 2 May 2016].
Faure, F., Corbaz, M., Baecher, H. and de Alencastro, L. (2012). Pollution due to plastics
and microplastics in Lake Geneva and in the Mediterranean Sea. Arch. Sci., [online] 65,
pp.157-164 & p582. Available at: https://infoscience.epfl.ch/record/186320 [Accessed 4 May
2016].
Hirai, H., Takada, H., Ogata, Y., Yamashita, R., Mizukawa, K., Saha, M., Kwan, C., Moore,
C., Gray, H., Laursen, D., Zettler, E., Farrington, J., Reddy, C., Peacock, E. and Ward, M.
(2011). Organic micropollutants in marine plastics debris from the open ocean and remote
and urban beaches. Marine Pollution Bulletin, 62(8), pp.1683-1692.
Monna, F., Dominik, J., Loizeau, J., Pardos, M. and Arpagaus, P. (1999). Origin and
Evolution of Pb in Sediments of Lake Geneva (SwitzerlandFrance). Establishing a Stable
Pb Record. Environmental Science & Technology, 33(17), pp.2850-2857.
NOAA Marine Debris Program, 2016. Marine Debris Monitoring and Assessment:
Recommendations for Monitoring Debris Trends in the Marine Environment. [online] Silver
Spring, MD 20910 USA, pp.24-25. Available at:
<https://marinedebris.noaa.gov/sites/default/files/Lippiatt%20et%20al%202013.pdf>
[Accessed 2 May 2016].
Plastic Debris in the Ocean. (2011). UNEP YEAR BOOK 2011, pp.2-13.
PlasticEurope, 2016. PlasticsEurope - Plastics the Facts 2013. [online] Plasticseurope.org.
Available at: <http://www.plasticseurope.org/Document/plastics-the-facts-2013.aspx>
[Accessed 24 Mar. 2016].
Queensland Government 2016, Department of Environment and Heritage Protection,
Ecosystem health indicators: Quality assurance and quality control, Brisbane, viewed 28
April 2016 via Griffith University database.
Reisser, J., Shaw, J., Wilcox, C., Hardesty, B., Proietti, M., Thums, M. and Pattiaratchi, C.
(2013). Marine Plastic Pollution in Waters around Australia: Characteristics, Concentrations,
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Seltenrich, N. (2015). New Link in the Food Chain? Marine Plastic Pollution and Seafood
Safety.Environ. Health Perspect., 123(2), pp.A34-A41.
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Two-way between-groups, in J Pallant (5th ed.), SPSS: Survival Manual: A step by step
guide to data analysis using IBM SPSS, Allen and Unwin, Sydney, pp. 274-284.
17
Warne, M., Batley, G., Braga, O., Chapman, J., Fox, D., Hickey, C., Stauber, J. and Van
Dam, R., 2013. Revisions to the derivation of the Australian and New Zealand guidelines for
toxicants in fresh and marine waters. Environmental Science and Pollution Research, 21(1),
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Lake Geneva Monitoring Program

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2203ENV Environmental Chemistry and Monitoring