Bio-investigations in the field

By Xavier Coadic

IN SHORT: An introduction to the concepts and practices of biological investigations (bio-investigations), including the collection and analysis of samples from the field and examples of research that you can do at home and in the local environment of your community.

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Our living environment is as much a book about the past and present as it is a permanent source of information. Whether it’s a hazardous chemical spill, a change in biodiversity, an urban renovation project or an improvement in water quality, we sometimes need to draw attention to clues and evidence from our physical environment, which often complement maps or other data we may find online.

Based on my (the author’s) field experience, this guide will present ways to explore, find and collect meaningful and “invisible” information that can be used in many investigations of our living environment.

To begin, all you need is a curious mindset and some methods to help you gather evidence of the existence of certain past or present environmental events.

What is a bio-investigation?

We call bio-investigation the processes of locating and taking samples and measurements in the environment, as well as preparing these proceedings and analysing samples or phenomena that will provide information, which can be incorporated into a broader investigation.

Bio-investigation, like other types of investigation, is also an act of creating knowledge so that it can be understood by others, by documenting the processes and actions that arise. Documentation will allow you to share your experiences and results until you are no longer the custodian of knowledge yourself.

For example, your bio-investigations can focus on polluted soils (soil, groundwater, soil gas), sludge and sediments (marine, estuarine, fluvial), air (ambient air, workplace air, air quality in a village or urban neighbourhood), etc.

This guide is dedicated to bio-investigation of the environment and it will cover both human and non-human activities.

Note:

The term “non-human,”, mentioned above, means “everything with which humans are in constant interaction:” animals and plants, but also elements such as water, air and earth, sometimes including objects and artefacts produced by human activity. This reminds us of other characteristics of these non-human elements involved in a situation.

For example, during the 2019-2020 bushfires in Australia, the media often focused on issues of disappearance of large animals, especially mammals, and the risks they faced. “Remarkable plants,” because they are aesthetically impressive, are also sometimes the subjects of emotional campaigns.

Microorganisms, whatever their reign, are usually invisible from public consideration, even though they represent a majority of the victims of these disasters and are thus a loss for the planet and humans in terms of the services they provide to the ecosystem.

Taking these “non-humans” into consideration of our vision of the environment during bio-investigations allows us to broaden our imagination, enrich our knowledge, grasp new sources of information, and consolidate our investigation process. Naturally, this consideration also allows us to take a more refined ecological approach to environmental situations.

A “common good” will then be considered as a “biotic community” bringing together humans and non-humans in a relationship of strong interdependence.

This guide proceeds in three stages:

• Describing why and how to organise yourself before conducting an investigation and taking samples in the field,

• Providing the basics of sampling procedures with different possibilities to analyse these samples,

• Showing you what you may find in different contexts with feedback from the field, and with real examples of connecting local issues with investigative and awareness-raising actions by groups of people on the ground.

It’s important to note that this guide refers to some basic steps to start using living material and your environment to disclose problems.

You will find methods, tools, tips and, above all, an investigative state of mind to be implemented in the ecological field, with environmental considerations concerning water, earth, air, plants, etc.

Investigate, cook, live

Studying our environment and trying “to make the living talk” is a bit like the process of cooking.

Learning to cook is something that one can start at a very young age and with little knowledge (and by enjoying eating well and making others enjoy the food), first by trusting the senses, and then by adding experience to the recipes. In bio-investigation, too, as we develop our techniques, we learn differently, we learn how to supply ourselves with ingredients, we acquire better tools and weave collaborative networks. Being an environmentalist or a bio-investigator is like cooking, perhaps by taking an interest in even smaller things/beings in life.

For example, a small scientific experiment to extract DNA from an organism may be considered filthy, unethical, complicated or risky. Yet you can do it yourself in less than 15 minutes in your kitchen on split peas or strawberries, simply with the products you probably have in your household. It’s fun, it demystifies a lot of methods and concerns, and it’s a first step towards discovering the invisible.

Photos from a workshop held in 2007 – “Biopanique cuisine et féminisme” (Biopanic cuisine and feminism) with DNA manipulation in a kitchen and a roundtable on women’s liberties. See the transcript of the workshop and the documentation of the DNA extraction process. Photos by Xavier Coadic.

Now let’s see how to get there.

Regaining control of information in your environment

People all over the world – in the countryside or in the city, in the mountains or by the sea – share a common goal: to gain an understanding of the information that concerns their surrounding environment and to participate in the further elaboration of this information.

This is especially true in the case of climate upheavals, accidents and natural or industrial disasters.

In June 2019, when an industrial abattoir spilled blood in one of the streams in Châteaubourg, France, or during the petrochemical industrial fire in Rouen in September of the same year, we (local citizen investigators and activists) found that many people were overwhelmed and that others wanted to simply meet over a coffee to discuss these disasters and to consider a new social configuration and organise themselves to address these issues.

So we, a local group of citizen investigators, gave as much importance to these circumstances of the meetings and the desire to work together as we did to the investigative methods and sampling protocols we wanted to deploy. It is almost certain that other disasters will occur, in other forms and under different circumstances. We have always adapted and, more importantly, have responded more effectively when we met regularly before or after an incident.

We have also experiencedt a re-appropriation of urban wastelands to limit gentrification and/or expropriations, with soils polluted by industrial heavy metals. We have had to deal with the pollution of coasts and beaches in Brittany by the oil spill or by chlorophytes, the decomposition of which releases a very toxic gas.

In each of these cases, it was necessary to deal with the emergency and regain control of the environment, the land and the information. Bio-investigation was an essential ingredient in this process.

Safety First!

Just as when you are cooking, gardening or tinkering, there is a golden rule in bio-investigations (and in any investigations): don’t put yourself or others in dangerous conditions. Here are some basic precautions:

Individual precautions:

• think before you act,

• prepare and plan your research. Below you’ll find some tips related to mnemonics, i.e. different constructions that facilitate memorization, which can be useful to keep long things in a short

• examine the place(s) where you want to take action,

• protective equipment is your friend,

• use appropriate equipment and training tools,

• don’t forget digital security tools and tactics,

• don’t work alone – even more so in the case of field (environmental) sampling,

• establish strong and trustworthy collaboration with others.

Collective protection:

• work in pairs or more as it will help and save you (e.g. climbing a wall or taking samples from a river),

• all group members should equip themselves carefully with all individual precautions,

• reuse, transform or develop methods and techniques for safe teamwork (drawing inspiration from urban graffiti, hiking, caving, diving, firefighting, etc.),

• document your work - you can get inspired from the experiences of common and resistance writing (or documentation as a form of resistance).

Remember that if one of you is in danger, the others will be in danger, too.

Operate in good local conditions

When creating, using and modifying the recipes, there are elements of culture, knowledge and experience that are shared and transmitted. This configuration is illustrated by the recipe of a Far Breton, for example – a typical cake from Brittany that says as much about the techniques and natural resources used to make it as about the customs and habits of the people who cook it. A cooking recipe, no matter how instructive it is, can be commented on by a considerable number of people in order to make adjustments as new (re)uses and adaptations are made.

This is the equivalent for environmental investigations: an area in which a simple commentary in the context itself can play a crucial role in following research methods and learning from them. So make sure you meet with the people who live on the sites you want to investigate and, if necessary, with their consent and after explaining the questions, interview them in a written, audio or video form (see “Interviews: the Human Element of Your Investigation”). You are certainly not alone in your concerns or your interest in this domain. Build relationships with indigenous people. The things you will do together are woven bonds that will leave connections and memories behind, creating a historical structure.

As in the situations of food crises, environmental tensions can be a theatre of powerful political debates and expressions of emotions. These crisis situations are the arenas of political and social discord that often lead to the dissemination of manipulated or even deliberately falsified information. Test, be vigilant and criticise these phenomena.

Example:

Here is a theoretical example of preparation before field sampling in the case of a factory fire that impacts water, air, soil, crops. Consider the weather and the extent of the disaster; identify and map potentially critical intervention and sampling points with an online mapping tool such as overpass turbo. Overpass turbo is a data filtering tool for OpenStreetMap – you can learn more in this OSM Wiki.

An industrial sulfur refinery site burns and two silos explode. The plant is located in a large city and in close proximity to a working-class neighbourhood with residential blocks and working-class houses.

This is the case of an industrial disaster – an industry that is central to the economic activity of a region and important for national consumption. The industrial site is maintained by an international company, and backed by associations and politicians who have been lobbying for this company for many years. On the other hand, there are politicians and associations that have been lobbying against these activities and this company.

During the industrial accident and in the immediate aftermath, many of these organisations could find it in their interest to disseminate or modify information in their favour. Add to this the concerns (and even panic) of the residents around the industrial site.

Prepare your field action plan and biological and chemical sampling scheme considering that the information that circulates and is produced during this accident may be profit-driven in one direction or another. Consider that the assistance you receive in the field while taking samples may influence the political interpretation of your investigation.

Protect yourself and know your limits

In order to carry out bio-investigations, you need to have detailed information about sampling authorisations: meaning, where you are allowed to collect samples from and where this is prohibited by law for various reasons. Biodiversity can be protected on the site you are targeting, or a health alert (e.g. risk of infection) may be announced. The relevant authorities, local associations and NGOs concerned very often disseminate this kind of information. You can also be guided by people authorised to carry out investigations in the field (scientists, associations, NGOs, academics).

In any case, act consciously and do not destroy or pollute nature.

Safety first!

When going to collect samples in the field, you must consider that you are usually observed by humans or devices (e.g. CCTV cameras). Even if you have the legal right to go there and nothing prevents you from taking samples, the fact that you are doing it may attract someone’s attention. Please think about it in advance.

Pay attention to video surveillance, monitor public or hidden camera locations, analyse the location in advance, walk the site before acting. In many cases, there are websites where you can view live images from surveillance cameras installed in the area, such as in Brussels, Seattle, or Rennes.

If you carry a smartphone or an IOT device, the geolocation and automatic WiFi connection can betray you and the other people who are next to you.

Apply good practices while in the field

Use the mnemonic allowing to make an inventory of the group and equipment. The key steps are grouped under the following list:

• Person(s) - Who are the people you work with in the field? What do you need to take care of these people? What are their needs and skills? Discuss together.

• Equipment - What equipment do you need to meet your operational objective? What equipment do you actually have? Check the availability, condition and functioning of this equipment.

• Workwear - What type of workwear is appropriate for your field operation? Check the condition of these outfits.

• Communication devices - What means of communication do you need in the field? Think about confidentiality and rules of constraint in case these communications are compromised. Make an inventory, test, train.

• Food - Will you need to hydrate and/or eat something once in the field? Provide food and drink for a set amount of time.

• Driving - Who does what? When? Where? Limit unnecessary effort and the risk of complications.

• Course of action - How will you proceed: objectives of an action, its duration, stages, planned replacements, etc.? Plan carefully.

• Appointments - Where, how, why, do you make an appointment to begin? Or when and why do you give yourself an appointment at the end?

Tips to help you move in groups or in pairs:

• Direction - From where are you departing, in what direction do you or the team need to go to reach the sampling site?

• Meeting point - What is the exact meeting point to start the work?

• Itinerary - Which intermediate points do you decide to pass through to reach the meeting point? Why?

• Training - In what configuration and with what means are you going to make the trip?

What you should remember once you get to the meeting point before launching the work:

• Situation - Description of the initial situation of the field in which you are going to intervene (e.g. a private agricultural field a few hundred metres from the factory, or within the perimeter of a surveilled building with demonstrations around it and police forces).

• Objective - What are your specific objectives to be achieved (e.g. 12 water samples at three different locations down the river + photos; or five air quality measurement points during 24 hours to form a study area in addition to some meteorological data).

• From idea to action - What are the techniques considered to achieve the objective? For each need, which precisely described techniques will you implement?

• Implementation - Task assignments and division of tasks / sectorisation. Make it clear who does what, how and where.

• Coordination - How you communicate, people to contact in case of problems, individual and collective security measures.

Theoretical example of preparatory work prior to taking samples:

On the river that was polluted last week, in the field of Mr. X who allows you to come on it if avoiding to damage any property, you have to take 12 samples from three locations that you have marked on your maps. The sampling techniques are described next to your maps. You have split into three teams of three people each. Each team has a designated person to supervise the sampling and security.

You have one hour to work on each location, including taking pictures. Communication between teams goes via Element/ex-Riot application with encryption enabled. Report any problems to the person supervising your team. Wear respiratory masks, latex gloves and do not come into direct contact with water to avoid contamination of samples. Take samples with a stick and small bottles that you then place in coolers.

Prepare your protocol for samples

Making a recipe

When you cook, you start with a need – to feed – and then a desire – to eat well. You prepare your idea in the kitchen, starting by making a list of the ingredients you have and/or what you need to get. Afterwards, you use tools to try to aggregate, gather, combine elements just before you start cooking, which is another affair in itself.

Drafting recipes of your sampling operations for a bio-investigation before going to the field will allow you to minimise the risk of forgetting details, to retrieve the sampling conditions, and to give a context of the samples (if you give your samples to other organisations for analysis).

Bio-investigation, just like cooking, is based on a recipe. You need recipes and other people probably need yours, too. So write down, document and share your recipe (suggestion: under free license); it will also be very useful to ensure the credibility of your investigation and its results.

Note:

Possible ways to share your investigative “recipes” with others (after testing and documenting them).

• A tutorial, which is focused on learning, allows the newcomer to take it as a lesson. It is similar to the process of teaching someone how to plant vegetables or cook.

• A goal-oriented form of a practical guide shows how to solve a specific problem and a series of step-by-step instructions. It is similar to the process of growing vegetables or a recipe in a cookbook.

• An explanation form, which focuses on comprehension and explanation and provides both background information and context. It is comparable to an article on the social history of the tomato or the cooking in general.

• A reference guide that describes a completed project. It is intended to be accurate and complete, thus verification and incorporation of sources are important. It could be similar to an encyclopedia article.

A good practice is to have trusted people read your sample collection protocol and ask them what they understand and what they don’t understand. This will improve the usability of your “recipes”.

For each measuring and sampling operation (physical, chemical, biological, meteorological, etc.), write a recipe beforehand.

Here you enter into the practices of metrology – through citizen investigation using measurement science.

Diagram made in a bar with people who wanted to practice bio-investigation. A pyramid of tile layers helping to symbolize the different areas to be chosen to define the places where biological and chemical materials should be collected. Diagram and photo by Xavier Coadic.

The inverted pyramid connected to the base pyramid by the red dot represents the continuation of the investigation process after the field operation. Diagram and photo by Xavier Coadic.

A bio-investigator’s mindset links the following actions:

• identifying/discovering methods and tools,

• inventorying sampling areas,

• investigating a particular situation,

• making field records and samples,

• analysing samples or measurements in this situation on defined and described areas,

• interpreting analyses,

• informing others (team, public, etc.) beyond your initiative.

Preparation scheme that represents the mindset to be adopted, as well as the methods and tools needed for field sampling. Diagram and photo by Xavier Coadic.

Building your protocol

When you examine your environment, the protocols matter.

Your protocols’ content will be influenced by the examples you find during your preparatory research, by the advice you receive, by the conditions in which you work, and sometimes even by the cases you pursue.

The important thing is to keep protocols simple to reproduce, to write them clearly, to test them, and then to adapt and improve them whenever circumstances require it.

Establish your own protocols as you would write a recipe. Whatever you do, you should always follow 5 procedures:

• geolocate your samples,

• measure and plot the sampling areas,

• take photos or videos of the steps you take,

• describe and comment on your work,

• make your protocols and procedures public if it doesn’t pose a risk (think about metadata and protection of individual data or anonymisation).

Tip:

Your recipe must also take into account the sequencing of your work, which is vital to the chain of custody and confidence in your samples. This is your logistics routine.

It’s like an envelope with a certification stamp that serves as a capsule on part of your sample’s life cycle (with its instructions). One capsule for each essential activity: sampling / measuring, storage, contextualisation, information indexing, transport.

The quality and integrity of these steps of your routine are important for the probabilities of reliable analysis and therefore strengthen your investigation (or weaken it if they are not sufficiently elaborated in the specific context).

Each routine is a specific treatment that must be well identified (object, nature, task, calculation, etc.) and tailor-made for each type of sample. A water sample will not have the same routine as a mud sample or a morphological imprint, even though the treatment may be similar (labelling for example). You can store your protocols in an online library (if your work is not confidential or putting you and others at risk) to make them available to other projects while preserving their integrity in their uses. For example, write everything down in a wiki, as if it were a recipe that you would like to convey with gestures, explanation of ingredients and associated data. It’s a process to be made visible and interpretable.

Diagram of the life cycle and stages of a bio-investigation with progression of the documentation and the amount of data indexed and made accessible. Diagram and photo by Xavier Coadic.

Bio-investigation routine schema and steps for documentation and data generation. Diagram and photo by Xavier Coadic.

To enhance the quality of your routine and associated documentation, you can use Ishikawa’s approach to cause-and-effect analysis using the cause-and-effect diagram or the following rule:

1. Material: materials used in the processes.

2. Equipment: equipment available and required.

3. Method: procedure to be followed.

4. Manpower: people available to carry out the process.

5. Environment: environment in which the process is carried out.

You may find this approach useful to control the quality of your routine.

Choosing sampling areas

You can use several methods and tools to select interesting and relevant locations to collect samples for your bio-investigation. Here we will give you new methods to identify areas for environmental information collection and physical sampling in the field.

Tip: Selecting sampling locations with Overpass-turbo

Overpass-turbo is a website that allows you to build maps with a few clicks by collecting data available in the OpenStreet Map databases. You can, if you have the skills, also use the code to build these queries, create a map and easily retrieve the associated data.

For example, if you have the ability to locate public drinking water points, or the surroundings of a school, or the location of a factory:

Click

Wizard`` / Assistant > search > school > build research > run

Or

Wizard / Assistant > search > amenity=drinking_water > build research > run

You will then obtain a standardised set such as: Codes + sources + data + maps.

Example of drinking water points. Source: https://overpass-turbo.eu/s/MG7

Schools. Source: https://overpass-turbo.eu/s/MG4

Factories. Source: https://overpass-turbo.eu/s/MKM

Here is documentation (in French) on how to prepare yourself for selecting sampling areas.

Energy-related infrastructures (oil, gas, electricity) with their facilities (production, storage, processing, transportation) represent a triple interest in your investigation:

• they allow you to consider a landscape according to its energy network,

• they allow you to know where to look for information in specific cases during your investigations,

• they have a considerable influence on the landscape and ecological environment.

Here is an example of a map, based on OpenStreetMap, accessible via a website with associated data. This particular map is zoomed on the city of Rennes in France:

Source: https://openinframap.org/#12/48.11283/-1.6717

There are many possibilities for taking samples and observing living organisms, some with simple protocols and basic materials, others with more elaborate methods. It’s like trying to make a dish cooked with similar but sometimes slightly different ingredients and changing the manufacturing process and cooking methods from time to time.

In this guide we provide some inspiring and proven examples that can serve as a basis for your ‘investigative mindset’. It’s also for training. But you will find many other possibilities and methods, more or less sophisticated, on the web. Don’t hesitate to look for them yourself, for instance by Dorking (see Search Smarter by Dorking in this Kit). Focus on the results contained in the free licensed wikis with notes about contributing communities. The best is to meet people with knowledge and experience in crafts (agriculture, gardening, fishing, DIY, biology, entomologist’s hobby, etc.) who could advise and help you especially in the beginning.

Illustrations of a 3-day introductory workshop on environmental investigation in 2017 with participants from a social integration programme. Photos by Xavier Coadic.

Taking soil samples

Creating a diagram of an area to be tested

There are many community gardens blooming in the city. Many abandoned areas are being reinvested in by communities, and developers are taking control of floodplains to build huge and expensive housing complexes.

Amateur gardeners occupying abandoned or fallow land are regularly confronted with the issue of soil pollution in these areas. Often, they are unaware of traces of bio-chemical inputs. In addition, soil analyses carried out in professional laboratories cost several hundred euro.

The invisible biodiversity of soils (fauna and flora, micro-organisms, plankton, bacteria, etc.) is almost never taken into account. It is estimated that several thousand species of bacteria and fungi can coexist in one gram of temperate forest soil. Each of these species is not represented by a single individual but by a multitude of individuals. In our gram of soil, we can count up to 1 billion bacteria and 1 million fungi and protozoa. Exposing them can help defend environmental causes and protect living conditions for all.

Here’s how to create a diagram of the experimental area:**

Source: http://aliceblogs.epfl.ch/years/y1_2018-19/studios/grandgirard/ferretti-lapo

• Make a diagram of the area where you want to conduct soil tests and indicate on the diagram where the samples were taken. For example, write “X1” for areas of high human activity (e.g., children often play there) or “X2” for areas of low activity.

Source : https://recaa.mmsh.univ-aix.fr/1/Documents/1-6.pdf

• Prepare your sampling plots in the field by dividing the land into several homogenous units/grids.

Source: https://mieux-se-connaitre.com/2011/02/larcheologie-sauvetage-de-memoire/

• Remove grass, stones, twigs, etc. from the surface of the ground you plan to dig.

• Using a spatula or spade, dig a hole 15 cm deep (you may need to dig deeper for some chemicals) and leave this piece of soil aside.

• Scrape the soil from the sides of the hole with a spoon (prefer a plastic or glass spoon, avoid metal).

• Collect about 28 grams (approx. 1 ounce) of soil all around the well.

• Every time you take a sample, take a picture.

• Every time you see an insect or plant, take a picture.

• Archive your samples with their associated note card (see an example here).

• Place the collected soil in a sealable plastic bag. Using a permanent marker, write on the bag the placement of the sample “X1” or “X2”, your name, the date of the test and the tests you want to perform (example: arsenic and lead). Seal the bag. Wash the spoon with soap and water, rinse with distilled water and use a clean spoon for each sample.

• Expand your assignment - Collect a total of 4 to 6 samples from the sectors in your study area, using the same protocol, each time with a clean instrument.

• When all samples have been collected, labeled and sealed, place them in a larger bag and store them in a container, along with the schematic of the investigated area that you have created.

• You can even give samples to the laboratory for analysis by an NGO, association or university if you have established a relationship of trust. Or analyse them yourself.

Once you have completed your searching, sampling, archiving and documentation work, compile everything into a file. This file can be then used to make a presentation of the area under study, including, for example, the discovery of high lead concentrations from former military cartridge mining activities on a site where a school is about to be constructed. Or, as another example, in the case of the construction of a marina complex, you could provide evidence of the existing biodiversity and its ecological importance that must be preserved.

Example:

You can also consider using art and science to spread the word, involve people, and build ecological action - such as making a Winogradsky column out of peat from an area of ecological interest on which a developer decides to start construction.

Example of a civic marathon initiated to design and test protocols for peat sampling in urban areas that are subject to real estate pressures. Photo by Xavier Coadic.

Photo by Xavier Coadic.

Photo by Xavier Coadic.

The associated map of the sampling (in green) and working (pins) areas in the city of Rennes in France. Source: https://umap.openstreetmap.fr/fr/map/byodit_151971#14/48.1128/-1.6762

Source: https://www.biointeractive.org/classroom-resources/poster-winogradsky-columnmicrobial-evolution-bottle

Source: https://schaechter.asmblog.org/schaechter/2018/08/how-to-build-a-giant-winogradsky-column.html

Warning: Note that investigating the soil requires taking precautions to avoid injury and to protect yourself from infections such as tetanus or yellow fever, for example.

Using plants as markers

Plants that grow in a defined area can help you imagine possible tracks of investigation regarding past events in that area. Indeed, plants, through the soil that filters water and provides minerals, absorb many elements from a past more or less close to the ecological life of the environment you wish to investigate.

Note:

Biodiversity concerns all living things, their interactions with each other and with their environment. The markers of this biodiversity are indications given by the presence in number and diversity of living organisms.

Start by identifying a site with a set of plants whose bio-indication and/or bio-accumulation potential you are looking for. You can then focus on mapping the location of these plants and collecting some specimens. This mapping, completed with the samples taken for analysis (at a laboratory or by yourself), can reveal valuable information on the state of the site and on the human activities that took place before these plants grew.

Numerous plants in your environment can serve as a reservoir of information when you are interested in a particular piece of land which, for example, is likely to have been exposed to heavy metals during human activities (factory, armed conflict, uncontrolled dumping ground). The presence of these plants does not necessarily mean that heavy metals are present in the soil. However, if you suspect that the soil on a piece of land contains heavy metals (lead, arsenic, cadmium, nickel, etc.), you may decide to collect plants with the capacity to absorb and store these metals in order to provide evidence after analysis. For example, ferns will be interesting to collect for your investigation if you have information that the ground on which they grow may have been polluted with arsenic.

Tip:

• See this Wikipedia entry of bioindicators for information only – it may depend on your ecological region and you must adapt the list by yourself.

• Also see this list of bioindicator plants (in French).

By searching on the internet or taking advice from farmers or NGOs specialising in plants as markers of biodiversity, you can more easily create a list of plants to be identified in the area of interest. Print out photos of these plants before going to the site for mapping and sampling.

It may be possible to follow a similar process by analysing the (micro)fauna in the soil, and the animals that eat it.

Note:

Plant samples degrade fairly quickly after collection. It is better to plan to collect the sample and get it to the laboratory (if you have this in the plan) on the same day.

Some rules of good practice:

• Wrap the samples in newsprint paper, which will block photosynthesis and absorb water vapor condensation, and package them in tightly sealed plastic bags.

• Store in a cold place (<10°C) for a maximum of 1 night before bringing them to the laboratory. If a ventilated oven is available, dry the samples for 24 hours at 70°C in suitable containers.

• Do not exceed this temperature, otherwise some of the volatile elements, especially nitrogen, may be lost.

• Repack in a plastic bag. The samples can then be stored for several months in a dry place.

• In any case, never close the bags with staples: a single staple is enough to contaminate the sample with iron and can damage the grinders.

The quantities to be taken depend on the analyses requested but also on the dry matter of a plant: a standard analysis requires at least 50g of dry matter to be able to grind the sample reliably.

Theoretical exercise:

Based on the list of plants given here, and their potential for indication or accumulation, try to make an exercise in a safe place (make sure of this before you start) near your house or town, for example in an abandoned construction site or industrial harbor.

Map this site locating these plants and then take plant samples for each type of plant. Finally, offer your samples to a soil and plant analysis laboratory (established by an association or a university near this site).

Measuring soil pH: an investigative exercise

Being able to determine soil qualities can be important in an environmental investigation. Tracking changes in soil qualities by conducting regular analyses, can help you understand changes in the environment. pH measurement describes the acidity (pH) of an environment. The hydrogen potential, noted pH, is a measure of the chemical activity of hydrogen ions in solution.

There are many tutorials available on the internet for soil pH analysis with digital sensors or test strips.

Some actions below require special tools if you want to implement them yourself at home. We provide examples of suppliers for such tools, but we do not endorse any particular vendor of these products.

The pH is a coefficient that characterises the acidity of a soil, due to the presence of H+ ions. It defines the concentration of H+ ions in the liquid state of the soil. The pH varies from 0 to 14 and neutrality is reached when the pH is equal to 7. Soils can be classified according to their acidity level as follows:

• pH < 4,5: highly acidic soils

• 4,5 < pH < 6: low-acid soils

• 6 < pH < 7: well-balanced soils for good mineral nutrition

• pH > 7: calcareous and/or saline soils

The pH is measured with a pH meter.

Protocol and work stages

1. Preparation of solutions

• Weigh exactly 10.0g of soil A, B and C.

• Pour each weight into a beaker which will be marked A, B and C.

• Add 50 ml of distilled water.

• Insert a magnetic stirrer into each beaker and stir the solutions for 30 min with the stirrer.

2. Filtration solutions

• Place a funnel on a high beaker.

• Place a filter (e.g., coffee filter) over the funnel and filter solution A.

• Do the same for solution B and then for C.

3. pH measurement

• Adjust the pH meter: calibration is done with a buffer solution at pH=7 and one at pH=4 (it is available in kits for aquariums or swimming pools in shops or via the Internet. You can buy buffer solutions for electronic pH meters - pH 4 and 7 online, for example from here).

• Insert the pH-meter probe into one of the solutions and read the pH value. Note the different pH values measured.

• Clean the pH meter and measure the pH value of the second solution.

pH meter. Photo by Sergei Golyshev (AFK during workdays), CC BY-SA2.0

Here’s an investigation exercise we suggest:

Investigators wanted to know where the soil found on the vehicle chassis came from., so they took three soil samples:

• sample A: from the location where the vehicle was parked.

• sample B: from the owner’s workplace.

• sample C: from the owner’s place of residence. Then they sent these samples to a laboratory.

Your task is to analyse these soil samples to determine the provenance/origin (as in location) of the soil found at the “crime scene.”

Note: Here we describe a study of the mineral salt content of a soil in order to identify the origin of the sample “A: from the location where the vehicle is parked”

Protocol and investigation exercise

To identify the presence of ions, tests involving a chemical reaction are necessary. To do this, a small amount of appropriate reagent (see table below) is poured into the test tube containing the test solution. Then you observe what happens and make notes about your observations.

You must have the following reagents: Sodium hydroxide (Na+; HO-); Sodium (2Na+; SO4²-); Silver nitrate (Ag+; NO3-) and Barium chloride (Ba+; Cl-).

Summary table of characteristic tests for ion identification :

Ion to identify	Reagents used	Observations
Fe2+	Sodium (Na+; HO-)
Fe3+	Sodium (Na+; HO-)
Ca2+	Sodium (2Na+; SO4²-)
Mg2+	Sodium (Na+; HO-)
Mn2+	Sodium (Na+; HO-)
Cl-	Silver nitrate (Ag+; NO3-)
SO42-	Barium chloride (Ba+; Cl-)

Pour approximately 2 ml of the sample containing the ion into a test tube using the pipette.

Then introduce a few drops of the reagent with a pipette. Observe and record your observations in the table. You will see colour changes, with or without precipitates (the formation of a dispersed phase in colloid in your test tube). These colours indicate a reaction that brings out a particular chemical component present in your soil sample, which will ultimately give important clues to the overall composition of a sample you are trying to trace.

Thus, 2 tubes with samples from 2 different sampling points with identical results and observations will give you an indication of high probability that the vehicle has been at both sampling locations.

Source: http://www.clg-moulin-lepecq.ac-versailles.fr/IMG/pdf/Chap_4_Tests_de_reconnaissance_de_quelques_ions.pdf. For ions Ca2+ producing a white precipitate https://webphysique.fr/test-identification-ion-calcium/

Download here a summary table of ion tests in pdf format.

This stage will help by your observations to determine the reaction for the presence of each ion (first column).

Determining the presence of ions in soil samples

• Pour 2ml of the sample to be tested into 7 test tubes to perform 7 tests as described in the table above.

• Run the ion determination tests.

• Write down your observations. Then run the same tests for samples B and C.

Comparison of your observations and conclusion

Using the previous results, you can now determine the composition and origin of the soil sample taken from the vehicle. Will sample B or sample C have the same reactions as observed by sample A?

The sample, among all those tested, which will show you the same results of reactions with identical colours or precipitates will also indicate the correspondence of composition with your soil sample taken from the vehicle. This will be important information that will help you trace the history of the vehicle’s movements. Like all important information, it will have to be treated with other contradictory processes.

Collecting samples for DNA analysis of the soil

Deoxyribonucleic acid, or DNA, is the carrier of genetic information present in all living organisms. It is specific for every species and may be unique to each individual of a species. Soil is the living environment for a large number of living species. From an ecological point of view, soil is one of the most important reservoirs of biological diversity on our planet.

One gram of soil can contain billions of bacteria belonging to several million species. Microorganisms have a fundamental role in soil function: organic matter dynamics, carbon and nitrogen cycles, degradation of organic pollutants, retention of metallic pollutants, etc. Microorganism communities also help measure all the environmental stresses affecting the soil. In this context, they appear to be good indicators of changes in soil quality and past soil-related events.

Soil samples collected as part of an investigation can provide meaningful and useful information following summary analyses, including for the genomes of microorganisms.

The protocol for soil sampling to assess soil microbial diversity is similar to that for physicochemical analysis described above.

Sample collection and packaging are critical steps that will ensure robust analyses and results on the characterisation of microorganism communities in the soil examples you have collected. As in cooking, the recipe and the actions are almost more important than the final dish. The objective of the protocols is to get from a few kilograms of soil from the sampling area a pill box (50 to 100 g) for preservation by ensuring that this “subsample” is representative. See more details and tutorials on the EcoFinders project website and pay a special attention to this video.

Note:

If you wish to go further in your investigations, you can download the protocol used for your extraction and then repeat this explanation of DNA sequencing (determining the genetic information of a living organism), or reuse this DNA precipitation documentation. If you become passionate about your work or you work on funded projects, you will find kits for advanced analysis on websites like the-odin.com from $99 to $1,900, as well as free materials and working methods on diybio.org, and a map of Do It Yourself Biology communities around the world that you can contact to build collaborations.

Source: https://sphere.diybio.org/

Protocol for taking and packing/storing samples

If you choose to send your samples to an NGO, laboratory or university for advanced analysis, here is a protocol that may help you. We will distinguish between 2 types of soil analyses:

• those to be carried out on fresh soil,

• those that can be carried out on dry soil.

Analysis of fresh soil: moisture at sampling, nitrates/ammonium, nitrogen and carbon mineralization test, microbial biomass, etc. The steps are as follows:

• Pack the soil sample in a hermetically sealed plastic bag (tie, knot).

• Label it with all necessary information.

• Put it in a cool (<10°C) and dark place as soon as possible (cooler then fridge) and transport within 2 to 3 days to the laboratory.

Analysis of dry soil: Here are the steps:

• Pack the soil sample in a thick, unsealed plastic bag.

• Put it in the shade, leaving the bag half-open in order to start drying, but avoiding the possibility of contamination of the samples (spilling the bag, animals or insects).

• Under these conditions the samples can be stored for 2 to 3 weeks before being sent to the laboratory.

If both types of analysis have to be carried out, the sample should be treated as on fresh soil, and drying will be carried out on a fraction of the sample by the laboratory itself. The quantities to be taken range from 1kg for a standard analysis to 10kg in the case of mineralization tests.

Note:

For 1 gram of soil, there are up to 1 billion bacteria and 1 million fungi and protozoa. See “Diversity of soil organisms” by Torsvick et al. (1994), Hawksworth (2001), Schaefer and Schauermann (1990).

These examples of working methods are also valid for many types of soil (mud, sand...). They enable – for example, in the case of a major factory fire with a plume of smoke travelling hundreds of kilometers – sampling to be carried out on several sites, over several months or years. Most importantly, these methods are easy enough for instructing others in group activities (just like cooking collectively).

Taking water samples

Soil receives and filters water from rain or human activities. The water then continues its cycle and we can observe it in several states: groundwater, rivers, marsh, sea and oceans. Just like plants or soils, different bodies of water can help reveal a lot of information about our environment.

Taking water from a river

Sterilize your container before sampling:

• in the oven at 170°C for 1 hour,

• in an autoclave at 120°C for 20 min, or

• in a pressure cooker with a little water for 40 minutes after the valve has been turned on.

Warning:

If you follow this procedure make sure you only use glass containers that can withstand this treatment. If you do not have this type of glassware, we also describe methods to take samples with plastic bottles. Keep in mind that the documentation of your protocol and precise information on the sampling conditions is more important than the attempts to sterilize the tools. A laboratory can process your samples if and only if you are rigorous on these points. If you perform analyses yourself, they will only be of value if you are rigorous on this point of documentation and precise information.

Carry your material packed in a chest cooler or a cooler bag to the sampling site in order to protect it.

For water samples taken from rivers, wells, etc. attach the sterile bottle to a rope, weight your device so that it submerges below the surface. Hold the bottle in the water with a pole to avoid your body being in contact with the water.

Water samples should be poured into sterile single-use bottles or vials (cleaned with disinfectant if necessary). Vials containing samples for analysis shouldn’t be completely filled – 3/4 full will be sufficient. Samples should be stored until analysis at 2° to 5 °C. If a chest cooler is used during field work, avoid exposing it to sunlight. Ensure the traceability of a sample (identification of the vial: date, time, title of your mission, initials of the persons, reference of the applied protocol, GPS coordinates of the sampling site). Samples should be analysed preferably within 8 hours of collection, otherwise within 24 hours.

Note: Here is an example of an aquatic sampling form.

In a notebook, write down everything you do and how you do it. You can also equip yourself with an aquarium test kit and conduct tests for pH, carbon dioxide, iron, etc. in the watercourse that will provide even greater precision to your operations. You can also use a thermometer and a Secchi disk.

Image of a Secchi disk. Source: https://en.wikipedia.org/wiki/Secchi_disk

If you do not have glass material for sampling, you can use plastic containersor sterile bottles that you can get from websites such as DeltaLab.

Tip:

In order to minimise the cost of acquiring materials and also to initiate meetings that could result in collaboration or help in your investigations, you can ask for donations of materials (bottles, glass containers, syringes, pipettes, pH kits, latex gloves, etc.) in pharmacies or a fire brigade or an infirmary near you. You will certainly find people ready to help you.

These institutions often have kits with professional equipment that are subject to very strict regulations and whose date of use has expired. The kits are packed in plastic to maintain sterility for a certain period of time. Laboratories are subject to very strict rules for the use of packaged kits. Even if professionals can no longer use them after the expiry date, you can do so if the packaging has not been damaged. Indeed, laboratories also carry out analysis of samples taken by citizens and stored in mineral water bottles. Though using kits that are still sterile but whose expiry date has passed makes the job simpler.

As with soil samples, you may consider using your water samples to detect the presence of dissolved metals. For a detailed guide, see the protocol established by the Ministry of Sustainable Development, Environment and the Fight Against the Climate Change (Québec, Canada). You can also watch this video on the technique of taking water samples from rivers or create a protocol for the waste inventory in rivers and on riverbanks, inspired by Riverine Input from the Surfrider Foundation Europe.

Safety First!

For your safety and that of the people you work with, remember that, for example, in the extreme cases that you suspect radioactivity in the water or a pollutant that causes high risk for humans, you must provide suitable, automated and secure means of collection.

Source : Water, Radioactivity and Environment. SFRP / Environnemental section, December 3-4, 2014. “Prélèvements d’eau dans l’environnement: de la théorie a la pratique”, Fabrice Leprieur, Benoît Philippot and all.:https://www.sfrp.asso.fr/medias/sfrp/documents/S4b-Fabrice_LEPRIEUR.pdf

Using river plants as markers

When taking water or plant samples from rivers or riverbanks, you must take precautions and have a clear methodology. Plants in these environments can be used for further tracks of your investigation or as evidence of the recent or distant past.

Safety First!

Be careful when taking samples in the field because some plants can also be dangerous for you (causing irritation / allergies). Such caution is valid in all situations and in all different areas (streets, fields, trees, rivers, etc.) during field examination and sampling. For example, Hydrocotyle ranunculoides, known commonly as floating pennywort, is an invasive water plant, potentially toxic to mammals. It contains saponins which sometimes causes foaming on the water surface.

Another non-toxic plant may also be of interest in your research – the river water-crowfoot (Ranunculus fluitans) aquatic herbarium, here in bloom (from June to mid-August) in the Dordogne region:

Photos by Xavier Coadic taken in Dordogne region in France

It reaches up to 6m in length.

River water-crowfoot is of ecological interest, it provides us with indications of the health and functioning of a river. The plant aids in purification: it assimilates parts of the nitrates and phosphates and oxygenates the water.

Ranunculus are homes for many insects (e.g. dragonflies), a refuge for juvenile fish, spawning places for some fish – a niche in which we can find other indicators concerning the river and its environment.

When ranunculus covers a river almost completely, it’s an indicator of:

• a state of no flooding for several years, which normally reshapes the river bottom,

• a low water level and high water temperature,

• the presence of nitrates in the water.

These indications provide you with potentially valuable insight about the river’s past and the environmental conditions and human activities in the river watershed – for example, pollution of the water by nitrates.

Example:

Here’s a project to show you how to collect elements from rivers – Hack The Panke:

• “Microbiologist Daniel Lammel and biochemist and illustrator Eliot Morrison from the artist and scientist collective DIY Hack the Panke will guide participants along the Panke River in Berlin-Wedding, discussing the interaction of water, soil and atmosphere from the molecular level and how this effects and is affected by organisms up to the larger ecological level. They will explain how water, plants and soil take part in cycles of carbon and nitrogen that support life, and how Berlin’s urban environment can both nurture and threaten biodiversity.”

In the case of river pollution in rural areas, try to organise yourself in third places by going to meet the farmers of the area concerned and people who could teach you new methods of soil, water or plant sampling. See the report about activities around a river that runs through the city of Rennes.

Water reserves and ponds

To take samples from reserves and ponds, apply the same methods and procedures as in the river. Remember that if the water reserve or the pond is on private property, you must obtain the prior agreement of the people who own the land.

In addition, you can find maps with data on groundwater quality on the web, for example here for France:

Source: Brgm // Infoterre http://infoterre.brgm.fr/viewer/MainTileForward.do and other websites with data for the UK and Australia: https://rivieres-pourpres.frama.wiki/documentation:outil:cartographie

Seas and oceans

If you want to take samples for seawater quality analysis, the bottles you use must be kept closed when they are not in use. They should not be washed with tap water but rinsed in situ with seawater before each sampling. This will avoid leaving traces of products or other elements from the tap water infrastructure in your seawater samples. The methodology itself is the same as described for rivers.

For the state of bathing water, the Surfrider Foundation has established a sampling protocol that you can use. The Surfrider Foundation also carries out bathing water analysis and regularly publishes the data obtained, as well as offering online courses to learn methods for monitoring the coastal marine environment.

Plankton

Plankton are microorganisms present in soil, river water or sea water. They are ecological indicators of the sampling environment. They are of interest to many citizens and university science programmes or eco-activist observation initiatives. The collection of plankton also represents many opportunities to collect other elements (micro-plastic, algae, shrimp...) and can result in collaborations that may be useful for your investigations.

Example:

The program implemented and documented at Concarneau allows you to acquire all the basics needed to bring together boaters, fishermen, citizens and scientists, and to manufacture plankton nets from curtains and PVC pipes. You will then be able to carry out investigations and participatory and citizen workshops by reusing the documentation, protocols and prototypes, as well as the collected data provided by programs that give access under free license.

Screenshots, source; https://www.notion.so/Programme-Objectif-Plancton-Concarneau-e367dc4bf5434766826c30ef9370da50

You can find comparable initiatives by searching online. This will allow you to compare methods and tools and to build partnerships.

Plastic waste on the coastline

During your bio-investigations you can map or ask for analysis of the waste that you find in rivers and on the coastline. With the same intention as sharing recipes, you can use other people’s tools, methods and data to feed your investigations and even to carry out your own bio-investigations, as well as by contributing to databases larger than the area you are interested in.

Tip:

Ocean Plastic Tracker is a web application for tracking and visualising geo-referenced waste reports:

Screenshot, source: https://oceanplastictracker.com/

You can combine your efforts in tracking and analysing plastic waste with those of your water microorganism sampling, water, soil and plant analysis. You will then be able to produce solid data and records to reveal pollution, inform a campaign to stop a construction project, support a collective project for site preservation, or even for use in your supply chain investigation.

Tip:

This is not a mandatory step in your investigation process, however, recording the sound during your observation and sampling operations in the chosen environment can help you contextualise your information in the future. It can even be used during the presentation, publication or exhibition of your research.

This requires time, skills, and specific equipment to record sounds – for example, around a watercourse or the park in which you are going to carry out investigations or in the heart of the industrial wasteland you are investigating.

It is up to each investigator or working group to assess whether this investment of resources and time makes sense in terms of the final objective of the investigation.

Monitoring air

There is a growing number of citizen science projects for monitoring and mapping air quality. This involves taking measurements of certain particles present in the air, rather than taking samples. These particles may come from industrial activities, pollination phenomena, vegetation fires, technological disasters or armed conflicts.

DIY tools and citizen actions with cartography

Here we examine the use of a DIY tool, Lufdaten, and some maps used for publishing air quality data, as well as examples of citizen workshops on air quality and metrology.

The electronic hardware and measurement protocols are well documented in several languages, so we will focus on mapping and especially on the possibilities of working in small investigation groups that can collaborate with each other on a larger scale.

Screenshot, source : https://luftdaten.info/fr/accueil/

Screenshot, source : https://luftdaten.info/fr/accueil/

AirBeam project – and its associated mobile application – may be of help for you.

For example, in 2018 in Rennes, France, citizens created the group Capteurs citoyens & qualité de l’air (”Citizen sensors and air quality”), meeting at Les Champs Libres in the framework of the RDV 4C (Créativité - Collaboration - Connaissances - Citoyenneté), every first Thursday of the month. Their action is based on public consultation but is carried out independently, with the Ambassad’Air operation (citizen initiative to control the air quality in the commune of Rennes alone). Check the documentation and reports of related workshops, in French only, and the AirBeam user’s guide.

Source: http://www.wiki-rennes.fr/Capteurs_Air_Beam

Example: micro-sensor data obtained by citizen investigators

The Luftdaten micro-sensors also show a high level of fine particles (PM2.5) in and around the city of Rennes in Brittany, France: Pacé, Vezin, Bruz, Servon.

December 4, 2019:

Source : luftdaten https://archive.is/saMD0

On January 17, 2020:

Source: https://rennes.maps.sensor.community/\#12/48.1031/-1.6867

Collecting additional samples

Measuring air quality from a fixed or mobile device can tell you about possible events that may have occurred at the time and place of measurement (such as a fire) or before measurement at a remote geographic location. A degassing of an industrial stack several miles away can be displayed on a sensor or on a map several minutes or hours after the event due to the influence of winds and other environmental parameters such as humidity, relief, etc.

Taking great care to protect yourself from accidents and contamination, do not hesitate to supplement these air measurements with samples taken as close as possible in the chronology of events with the methods described above. You will be able to complement these samples with collections of soot, sludge, ashes, oil pellets (that you can for example scrape with a knife from a street lamp or a house roof). Place all this in clean and waterproof containers and remember to include descriptive notes. Keep the mindset that we try to describe here.

Safety First!

Whether you work alone or in a group, going to investigate your environment and taking samples will make you an object of curiosity with the public, including those who do not share your ideas and intentions. You will attract attention. Therefore, try to remain discreet.

In addition, the simple act of transportation of samples, biology and chemistry equipment may attract police or private security forces. In France, for example, since 2015 and the generalisation of the state of emergency after the terrorist attacks, police services have been taking more and more liberty to search, confiscate and destroy materials for increasingly vague reasons. Carrying your tools and equipment in a backpack can result in a police check and the immediate seizure or destruction of your equipment. At train stations and airports this can go even further, potentially resulting in an arrest.

In many parts of the world the simple act of putting a sticker on your laptop or sewing a patch on your bag can attract the attention of the police or private security services. In some countries, using the Tor browser or the Tails operating system*** can make you a target for prosecution.

In this kit, you’ll find safety recommendations in each guide as well as a stand-alone introductory Safety First! Guide to help you assess your risks and prepare to mitigate them. You even have a guide to learn how to behave in the field: Away from Your Screen, Out in the Field.

This kind of ostensible labelling can get you in trouble. Source: Xavier Coadic.

You may meet people, including police officers, who label you an “eco-terrorist.” It will take a great deal of patience and fortitude to explain that your actions have nothing to do with any form of terrorism. It is difficult to prepare for this, but you have to be very serious about it. You also have to think about protecting those around you from the repercussions of such allegations.

In late October 2019 in France, a biologist and professor investigating the effects of tear gas used by the police in response to social movements was arrested and held in police custody for 48 hours, and his home was searched while he was in detention. On his personal Facebook account, the professor listed the items that were searched at his home and in his vehicle. These included his laptop PC, USB sticks containing his lectures and his work on tear gas, several books on the police and the history of tear gas, and a number of empty tear gas grenades.

Within the framework of your bio-investigation, you will certainly have to return several times to a site to carry out observations, sampling and measurements, sometimes over several months or even several years. This repetition of bio-investigation activities may reinforce curiosity and the interest that a particular population or organisation may have for you.

Other markers and data in your environment

To complete your bio-investigations you can use other elements of the landscape that are symbolic for a wider audience, rather than just the target group that might be interested in the subject of your investigation.

For example, in the case of a shopping mall construction project, you might take a remarkable tree as an iconic totem of your bio-investigative work. This can provide both opportunities to collect a lot of data that will be useful in your investigation and to mediate your work and the ecological environment.

Sketch illustrating the processes of bio-investigation work on and around a tree selected as an iconic totem. Source: Xavier Coadic

Taking this illustration as an example, draw your own plan with a grid, as described earlier in this chapter. Indicate the GPS coordinates on your sketch and the North/South orientation. Note what tree it is if you know, or add a detailed description, the circumference, the GPS location and its proximity to a building or other remarkable construction. Take photos of the trunk, leaves, surrounding soil and insects you find.

Get sample note sheets similar to the example provided in this chapter. Take samples of soil, water if there is water on the ground, plants present near the tree, and the leaves of the tree. Fill in your sampling sheets as you go along. Annotate and carefully package all your samples.

If present, identify mosses and lichens on the surface of the tree trunk.

Source: University Paris Sorbonne: https://www.particitae.upmc.fr/_resources/doc/

Lichens as markers

Lichens are present everywhere in our environment: in the mountains, by the sea, in the city, on stone, on wood, on metal. Also lichenised fungi are composite organisms resulting from a symbiosis between at least one heterotrophic fungus and microscopic cells possessing chlorophyll (green algae or autotrophic cyanobacterium for carbon).

Lichens grow very slowly. On average, annual growth is between 0.5 and 2mm for crustose lichens, 0.5 to 4 mm for foliose lichens and 1.5 to 5mm for fruticose lichens.

They can be excellent markers of the health of the environment in which they are found. It is not mandatory to be a lichen specialist, nor to know how to identify them perfectly to carry out an inventory and take samples for bio-investigation. We’ll give you some inspiration to get you started.

Note:

Specialists distinguish several types of lichens according to the overall appearance of their thallus (the vegetative apparatus of a plant, devoid of conducting vessels and therefore forming neither roots, stems nor leaves in the strict sense). Here is a list of lichen types from Wikipedia:

Common groupings of lichen thallus growth forms are:

• fruticose – growing like a tuft or multiple-branched leafless mini-shrub, upright or hanging down, 3-dimensional branches with nearly round cross section (terete) or flattened.

• foliose – growing in 2-dimensional, flat, leaf-like lobes.

• crustose – crust-like, adhering tightly to a surface (substrate) like a thick coat of paint.

• squamulose – formed of small leaf-like scales crustose below but free at the tips.

• leprose – powdery.

• gelatinous – jelly-like.

• filamentous – stringy or like matted hair.

• byssoid – wispy, like teased wool.

• structureless.

For the purposes of your investigations, we focus on three main types of lichens based on three morphological distinctions of the thallus:

• Crustose lichens: that you can mark with a C in the inventory sheets,

• Foliose lichen: that you can mark with an F,

• Fruticose lichens: that you can mark as Fr.

What you will need:

• a camera,

• a device providing GPS coordinates (for example, a mobile phone with GPS),

• a small magnifying glass, ×30 minimum,

• curiosity,

• something to take notes on,

• a ruler,

• a piece of wire mesh or something similar with dimensions of one square marked.

Example of crustose lichen on a fence near a crematorium. Source: Xavier Coadic

Fruticose lichens are rather sensitive to air pollution and are therefore rare in cities. Equipped with your cameras, sampling kits and inventory sheets, you can take observation notes such as ground heights, North/South orientation, photos with the magnifying glass. Repeat this from time to time, marking pollution events or major disasters that occurred just before your last investigation.

Example with a crustose lichen on a tree. Make an inventory and locate the lichens at the seaside by using a piece of plastic waste on which the diameter of the circle is indicated with a felt marker, with the 4 cardinal directions correctly oriented. Photo: Xavier Coadic. Sentier des douaniers. Cancale

You can then multiply the number of locations in town, design a rigorous inventory file with all the contextual information (GPS location, height on the ground, weather, date, etc.) and make a file or complete the work scheme around the tree as described before without being a lichen expert.

Example of fruticose lichen (in yellow and orange) on a branch of a bush by the sea in Brittany. Photo by Xavier Coadic

Example of foliose lichen on a rock by the sea in Brittany. Xanthoria parietina, mushrooms and algae symbiosis, yellow thallus and orange apothecia. Parmelia of the walls. Photo by Xavier Coadic

Don’t be afraid to make a mistake. Just like in cooking, you learn by practicing and understand by documenting.

Note:

Some organisms known for accumulating pollutants are or could be used as bioindicators or for environmental assessment (biomonitoring). For example:

• lichens, by accumulating the pollutants, enable a retrospective analysis of their exposure to heavy metals or radionuclides ;

• the zebra mussel (freshwater mussel, dreissena polymorpha) accumulates metallic elements in the canals;

• fungi also have potential to biomonitoring, especially for metals (Source: Wikipedia).

Flowers and yeasts

In your direct environment, you may also find surprising opportunities to conduct parts of your bio-investigations, while having the freedom to collaborate on other science and ecology projects at the same time. This means you can gather even more interesting and reliable information and data while continuing to learn more and sometimes getting even more surprised.

In spring and summer, there are probably flowers in trees and bushes all around your place. Be aware that many of these flowers are naturally covered with yeast, which is of great interest to scientists, bakers, amateur or professional beer makers. These flowers also contain a lot of other information on the quality of the environment, on genetics, on biodiversity, and on the plant heritage of the region. So much information and data that can easily be incorporated into your investigations if needed.

This example of BrewLab explains how to capture wild yeasts, how to cultivate them, how to obtain or produce your material and even proposes to go further with the precise identification of these yeasts by genetic analysis. Although BrewLab researchers are experimenting with new types of beer from wild plants, their research and tips are a useful learning resource for citizen-led bio-investigations. You will also find a wiki page to guide you in this adventure. Or do similar experiments with potatoes, which are very common in everyday food consumption and agriculture.

Products you consume

Investigating the ingredients present in mass-market products is possible with OpenFoodFact, a mobile application that uses barcodes to give you access to free databases.

Source: https://fr.openfoodfacts.org/decouvrir

This information and data can help you in your bio-investigations, especially if you can determine the manufacturing or farming locations - part of the supply chains - of these products or their ingredients and then use the sampling and measurement protocols from this guide. Learn more about researching products and their supply chains from the chapter Supply Chain and Product Investigations in this Kit.

Analyse or share your samples

We have parts of ourselves to retrieve, understand and react to while investigating our environment. By conducting bio-investigations, we collect both Bits (information units) and Bits (pieces) of Life.

This is what you will see during the meetings that will mark out your investigations in the field.

The analyses of the samples you have taken, the measurements and the observations you make during your field actions within the broader framework of the theme of your investigation contribute to this reconstruction of a narrative and the re-appropriation of parts of life.

It’s up to you to choose whether to outsource the analysis of the pieces of information, to whom and under what conditions.

Share your samples for analysis by others

Once your field activities are completed, you will need data to be extracted from your samples, your measurements to be analysed, and then followed by data qualification and interpretation. This is a very important stage to transform your efforts into a usable material relevant to your investigation purpose.

We have mentioned situations where other organisations potentially interested in your work might have different interests from yours and therefore use this data to serve a completely different purpose than your own.

When choosing a third party to conduct a part of the analysis you need, you should keep these risks in mind.

You can choose to work with hackerspaces or biohackerspaces (a list is for example available here), or NGOs or other associations that have the analysis and processing capacities specific to your topic and your samples, or even university and state labs or private companies.

Not all laboratories or community facilities have the same analytical capacity and expertise. Your choices will also be driven by these possibilities or impossibilities.

These tests can be free or paid, and sometimes very expensive. The high cost of analysis should never be used as a way to blackmail you. If this happens, do not collaborate with the organisation that is doing this to you.

In any case, try to be accompanied by a legal advisor, establish a clear collaboration contract, whatever form it takes, containing your rights and duties, the price or the explicit exemption from payment of the tests, the rights and duties of your working partner(s) and above all the clear conditions of confidentiality and exploitation of the data that will be derived from the analysis.

You are never required to give all of your notes, observations, samples and measurements to a single organisation. You can even distribute them among several organisations in order to have material to compare and also to keep a part of it to carry out similar or different analysis yourself.

Bio-investigation as a community act

It may seem long and fastidious to prepare and then carry out a bio-investigation, but it is a process that gives you a lot of satisfaction as you advance in your methods and learning. Just like cooking.

You can, for example, set up biology cafés in a bar in the neighbourhood that is subject to real estate pressure to teach people how to organise themselves in small working groups. You can then discover, in collaboration with people concerned by environmental issues, other methods of working and sampling. See this example from the author – “Ateliers de Biologie” in Rennes.

Bio-investigation and its methods can easily become a mindset and a community action, especially when you need to act to change an existing situation. Bio-investigation allows you to learn more about your local environment and the possible problems it confronts. These are your issues, as well as of your community. Get involved.

Acknowledgements:

This guide would not be possible without meetings, collaborations, mutual aid and trust. Although it is written by one individual, it is the result of work of many people who have contributed to it in one way or another. Thank you to: Mily1000V, Laura, Wael, Hackorn, Emmanuel Poisson-Quinton, Jacques Le letty, Rieul Techer, Yann Heurtaux, Asso Ping à Nantes, Ewen, Jaxom, Sylvia Fredriksson, Le Biome HackLab in Rennes, La Myne in Lyon. Thank you to Sasha Gubskaya for the English translation and adaptation of the initial French-language guide.

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Published July 2020 in French, and December 2020 in English

Resources

Articles and Guides

In English

• Alabama Environmental Investigation and Remediation Guidance from The Alabama Department of Environmental Management.

• DNA Sequencing from Open Food Repo DNA.

• Hazardous materials investigation, sampling, & documentation from The Washington State Department of Transportation.

• Lichens Biology, guides and tips from the U.S. Forest Service.

• Successful Microbiological Investigations, by Scott Sutton.

• Archeograph.com. A collection of tutorials, graphic and cartographic resources for graphic designers and archaeologists.

In French

• Accueillir les Non-Humains dans les Communs (Introduction) by Lionel Maurel.

• Analyse du sol à faire soi-même. A low-cost and reliable soil analysis kit for gardeners. From PING association in Nantes, available on Fablabo.

• Capter et (co)produire des savoirs sous contraintes : le tournant expert de Surfrider Foundation Europe by Julien Weisbein.

• Comment composer avec le monde « non-humain »? by Philippe Descola. France Culture, broadcasted on January 3, 2015.

• Échantillonnage et analyse des sols pollués. Analysis framework from the Institut de veille sanitaire, France.

• Guide pratique pour la description des sols en France from the Conservatoire d’espaces naturels de Bourgogne, France.

• Guide d’échantillonnage à des fins d’analyses environnementales from the Centre d’expertise en analyse environnementale du Québec. (archived copy from Wayback Machine available here.)

• Lichen, Agent de la qualité de l’air by Mathias Roth.

• Manuel des Protocoles d’Échantillonnage pour l’Analyse de la Qualité de l’Eau au Canada from the Canadian Council of Ministers of the Environment, 2011.

• Méthodologie de diagnostic des sols pollués by Hafid Baroudi (workshop entitled «polluted soils», june 1997, Paris, France. pp.3-7. ffineris-00972099f.)

• Protocole d’échantillonnage de la qualité de l’eau from the Ministry of Sustainable Development, Environment, and Fight Against Climate Change (MELCC) and the Conseil régional de l’environnement des Laurentides (CRE Laurentides), 2017.

• Reconnaître et caractériser les zones de forge sur surface décapée. Apport de la géophysique à l’étude paléométallurgique by G. Hulin, B. Jagou, M. De Muylder et al.

• Recette frugale d’hackathon citoyen open source: en 32 jours e sans budget.

• Stratégie et technique d’échantillonnage des sols pour l’évaluation des pollutions by L. Belkessam, B. Lemiere – CNRSSP.

Tools and Databases

• OpenStreet Map. A free open source map platform created through crowdsourcing.

• Overpass-turbo. A website which enables you to build maps with a few clicks by retrieving data available in OpenStreet Map databases.

Glossary

term-beaker

Beaker – a container used for many actions in laboratories, especially in the domains of chemistry, physics, biology and pharmacy (https://en.wikipedia.org/wiki/Beaker_(laboratory_equipment.)

term-bioindication

Bioindication - Information provided by a plant, fungus or animal species, or by a group of living organisms about the state of the environment and/or its past, including the impact of certain human practices on it.

term-barcode

Barcode – a set of symbols (series of small lines / bars) applied to consumer products to identify the manufacturer, as well as additional information on the product / its origin.

term-bioaccumulation

Bioaccumulation – some organisms’ (plants, animals, fungi, microbes) capacity to absorb and concentrate in their organisms or in a part of their organisms (living or inert part such as tree bark or wood, mussel’s shell, horn, etc.) certain chemical substances that are possibly rare in the environment. This is the principle of bioaccumulation.

term-biocommunity

Biotic community – also called ‘biota’ or ‘biocoenosis’, includes all living and non-living (rocks, ashes, tools) creatures that live interdependently in the same delimitable area (see more here.)

term-biodiversity

Biodiversity – a variety of life forms on Earth. It can also be considered as the variety of living organisms present in a more specific area.

term-commons

Commons (or common goods) [Elinor Ostrom] – a notion that applies in domains of, for example, the nature management and in a number of other fields (Knowledge Commons, Digital Commons, Social Commons, Urban Commons, etc.). A common good is an actually existing good around which people come together in conviviality to maintain this object under shared responsibility and with deliberate rules of governance.

term-distilledwater

Distilled Water – a generic term covering several methods of purifying water to remove, for example, mineral salts or biological substances.

term-dna

DNA – deoxyribonucleic acid (DNA) is a biological macromolecule present in all cells of living organisms, also in some viruses. DNA contains all the genetic information, known as the genome, that enables the development, functioning and reproduction of living beings. DNA is composed of nucleotides (CGAT.)

term-dnaseq

DNA sequencing – the process of determining the sequence of nucleotides in a DNA fragment. The reading of this obtained sequence allows to study the biological information contained in it.

term-genome

Genome – the genetic material of an organism.

term-grid

Grid – a geographic/topographic drawing that accurately divides the physical area of sampling to reference each square by size, abscissa and ordinate (see https://en.wikipedia.org/wiki/Abscissa_and_ordinate.)

term-gridmap

Mapping the grid – integration of a grid into a geographical map, with topography if necessary.

term-totem

Iconic Totem – a symbolic representation that serves as a demonstrative emblem aimed at making low-visibility phenomena recognizable.

term-iot

IOT – Internet Of Things, refers to a connected object which by a given protocol allows to operate in a larger network.

term-metrology

Metrology – a measurement technique applied to a particular field.

term-mnemotech

Mnemonic devices – methods and tools helping to facilitate the memorization of information.

term-morphoimprint

Morphological imprint – a characteristic specific to the shape and structure of a body (human face or paw prints, fossils).

term-oligoelements

Oligo-elements (trace elements) – mineral substance, which our body needs to function. They are present in water, soil, plants and animals.

term-pHmeasurement

pH Measurement – measurement of the acidity (pH) of a medium. The hydrogen potential, noted pH, is a measure of the chemical activity of hydrogen ions

term-photosynthesis

Photosynthesis – a biochemical reaction of energy creation directly from mineral compounds using the light energy provided by the sun. The reaction takes place in living organisms (plants, algae, bacteria, etc.)and is necessary for the organism’s life.

term-routine

Routine – a set of precise and repetitive rules implemented over a period of time within an activity.

term-sample

Sample – in bio-investigation, fragments or elements of objects in a defined area (sounds in a recording, leaves on a tree, the waters of a river). A sample is representative for the population or the area from which it was taken.

term-sampling

Sampling – actions of taking and classifying samples of objects from a defined area (sounds in a recording, leaves on a tree, water in a river).

term-supplychain

Supply chain – a set of stages that commodities (goods or raw materials) must go through before they become products used by consumers or industry.