Citizen Science at the Ontario Wolf Survey

We equip interested members of the public with DNA capture kits to help us collect non-invasive samples across suspected Algonquin wolf range. Please contact us if you are keen to learn more.  DNA can be collected on private property with permission, and on crown land. DNA samples can be collected in provincial parks provided there is a research permit in place. It is illegal to collect in parks or other protected areas without a research permit.

This citizen science surveys expand our sampling efforts, and provide critical data to the province’s monitoring and recovery of the Algonquin wolf. Collecting DNA non-invasively entails looking for urine/scat and hair samples and storing them frozen or swabbing DNA from scat in the field under certain circumstances. Citizen scientists must first read safety information (found below), be physically able to walk trails, and able to store DNA samples in their own freezers until samples can be submitted (typically once per year at end of spring). Samples are small and the average citizen scientist will collect less than 10 in a full season.

How to Find DNA Samples

Sample Collection Instructions

Wild canids (wolves, coyotes and foxes) are territorial species, and they mark their territories using urine and scat. Since our sense of smell is not nearly as well developed, we rely on snow to be able to track wolves/coyotes and locate their urine marks and scats. Snow also helps keep the DNA intact for lab work.

Like people, wild canids prefer to walk along trails and roads to save energy, especially in winter when snow is deep. Since they want their markings to be easily noticed by other canids to communicate territory boundaries, they often urinate or defecate in noticeable areas.

Look for urine on the side of snowbanks, especially at the intersection of a trail or two roadways, or against the base of a prominent feature on the landscape (a big tree in the woods or a boulder of snow that falls back on to the road after the road has been plowed).  If a urine sample smells skunky, it is a fox urine, and should not be collected. Do not collect urine samples in areas where dogs frequent.

Look for scat in the middle or on the side of the road. Sometimes a canid will jump on top of the snowbank and defecate there, or walk along a trail for some time and deposit scat at the top of a slope or on top of a big rock. Generally, wolves do not deposit scat in the middle of the woods, but they may defecate near bodies of water, such as shorelines of a big lake, or on a land bridge between two water bodies. Wild canids and domestic dogs eat different things and usually their scat looks noticeably different, so you can collect scats even in areas where dogs might be present.

Wolf and coyote tracks are bigger and more clearly defined than fox tracks, which often look fuzzy and do not sink deeply into snow. Wolves and coyotes can have tracks that look similar to medium-to -large dogs. However, dogs often travel in a zig-zag pattern, whereas wild canids usually walk in a very straight line, diverting only sometimes to investigate things on a trail. Wolves and coyotes have narrow chests and their tracks can look like a single line of individual paw prints; this pattern is called “direct register” because the rear foot falls directly on the spot where the front foot stepped. Dogs do not often have perfect direct register, being less efficient, “sloppy” walkers. Identifying wolf/coyote tracks apart from dog tracks can be difficult, but remember that dogs are usually accompanied by parallel set of human tracks!

 

Two tracks intersection: smaller, fuzzier looking fox tracks (horizontal), and larger wolf/coyote tracks (vertical).

Two tracks intersection: smaller, fuzzier looking fox tracks (horizontal), and larger wolf/coyote tracks (vertical).

Before sampling, you must read and abide by the following safety information sheet:

Echinococcus info sheet

 

Where is the DNA?

DNA is found in bladder cells sloughed off in urine. These cells are not visible to the naked eye.

DNA is found in sloughed-off intestinal cells found in the mucus that surrounds scat. Mucus is not always visible, especially on older scat, but it looks like a shiny, clear layer when visible. It is often visible around the terminal “pinch points” on the scat.

 

How are samples collected?

Urine samples are collected into 50mL tubes with screw top lids. If the sample is large and there is more urine-saturated snow than can fit into the tube,  try to collect the most concentrated (darkest yellow) portion of the sample. Unique sample ID codes are written with permanent marker on all tubes. Samples are to be kept frozen.

Frozen scats are collected whole into freezer bags with unique sample ID codes written on the bag with permanent marker. The scats are later swabbed in the lab to remove the DNA.

Thawed scats are sampled in the field by swabbing the external mucus layer with a sterile swab that looks like a big Q-tip. Look for visible mucus, often a clear sheen, and roll the swab head over it to roll up the mucus. To deposit the DNA from the swab head into the liquid buffer solution in the 2mL screw-top tube, vigorously swish and press the swab head against the inside of the tube to loosen the DNA. Then, swab the rest of the scat. The swab should be swished in the liquid in the tube several times throughout swabbing. Do not pierce the scat with the swab or collect hair from the scat, otherwise the DNA results will show the prey species that the canid consumed (e.g. beaver or deer) rather than the canid itself.

Even if swabbing the DNA from thawed scats in the field, collecting the swabbed scat into a ziploc bag is important if diet analyses are being undertaken. If in doubt, collect the scat. If you do, make sure both the scat bag and the 2mL sample tube are marked with identical sample ID codes so that we know they go together. This is also useful if insufficient quantity of DNA is found in the 2mL tube, because the scat can be swabbed a second time in the lab. Freeze the scat sample as soon as possible.

 

Please download the full DNA collection instructions found below. These instructions come laminated in DNA sampling kits. If you are interested in obtaining a kit, please contact us.

DNA capture instructions

Large canid tracks. Sometimes the front and hind tracks do not overlap, which shows the size difference between them (front paws are larger).

Large canid tracks. Sometimes the front and hind tracks do not overlap, which shows the size difference between them (front paws are larger).

Large urine sample found on the side of a snowbank.

Large urine sample found on the side of a snowbank. Such large samples are typical sign of dogs rather than wild canids, however urine samples like this can be collected if you are travelling in an area far from homes and human tracks.

Measurement of large canid tracks. Note the front and hind track overlap here, which is called "direct register", making the animal's trail look like a single tracks in single-file.

Measurement of large canid tracks. Note the front and hind track overlap here, which is called “direct register”, making the animal’s trail look like a single tracks in single-file.

Small urine sample found along beside wolf tracks. Note small size against 30cm ruler.

Small urine sample found along beside wolf tracks. Note small size against 30cm ruler.

Large canid diverted from walking along he middle of a trail to urinate on a small tree at the side of the trail.

Large canid diverted from walking along he middle of a trail to urinate on a small tree at the side of the trail.

In late winter, female wolves enter into annual estrus cycle, and blood spots can sometimes colour urine sampled a dark orange or red.

In late winter, female wolves enter into estrus, and the resulting blood spots can sometimes colour urine a dark orange or red. Blood-saturated snow at kill sites might be from the prey or less likely, from an injured canid. Do not disturb fresh kill sites, an leave the site, but note its location and report it to us. Blood-saturated snow should be collected into 50mL as you would for urine, and kept frozen. Make notes about what you see to help us determine whether the blood is from a canid or its prey.

Scratch marks in front of a scat sample found in the middle of the road.

Scratch marks in front of a scat sample found in the middle of the road.

Very fresh scat (A on freshness scale). Fresh scats look wet and shiny, even if frozen.

Very fresh scat (Freshness scale A). Fresh scats look wet and shiny, even if frozen.

Scats are often deposited in the middle of the road. Only collect road scats if you can do so safely, obeying all signage.

Scats are often deposited in the middle of the road. Only collect road scats if you can do so safely, obeying all signage.

Large canid scat showing two different fecal matter types: hair (typical) and digested flesh/organs from prey (dark brown). Scats that look like twists of hair almost always belong to a wolf/coyote rather than a domestic dog. Dog scat can have a few hairs in it from self-grooming.

Large canid scat showing two different fecal matter types: hair (typical) and digested flesh/organs from prey (dark brown). Freshness scale: B. Scat that looks like twists of hair and bone fragments is almost always from a wolf/coyote rather than a domestic dog. Dog scat can have a few hairs in it from self-grooming.

Large canids often divert from trails to hunt along shorelines. Do not follow tracks that lead to water bodies for safety reasons.

Large canids often divert from trails to hunt along shorelines. Do not follow tracks that lead to water bodies for safety reasons. However, you may find scat on the shoreline.

Unusual canid scat showing hair twist on one terminus, and the other contains vegetation.

Unusual canid scat showing hair at one terminus, and the other terminus is made up of undigested vegetation. This is an older scat, the hairy terminus is no longer smooth. (Freshness scale: C)

A large canid walking along a roadway diverted to mark the snowbank with both urine and scat. Count the number of tracks to better estimate whether the samples come from one individual or more than one. Often, an older scat or urine mark will be later marked a second time by a passing canid.

A large canid walking along a roadway diverted to mark the snowbank with both urine and scat. Note the long scratch marks to the right of the samples. When you find more than 1 sample together, count the number of tracks to better estimate whether the samples come from one individual or more than one. Often, an older scat or urine mark will be later marked a second time by a passing canid.

Note the frozen, clear mucosal film along top edge of the largest section. Mucus is brittle when frozen.

Note the frozen, clear mucosal film along top edge of the largest section. Mucus is brittle when frozen. This scat is older (Freshness scale B) but the mucus is in good condition, making it a good sampling candidate.

If you not sure how to identify tracks to species, take a photo of the track with a ruler or other known item to show reference size so that it can be measured later. Left side track is the smaller hind paw, right side track is the larger front paw.

If you not sure how to identify tracks to species, take a photo of the track with a ruler or other known item to show reference size so that it can be measured later. Left side track is the smaller hind paw, right side track is the larger front paw.

Old scats look furry if they contain hair, rather than smooth. If they do not contain hair, they look powdery and white/grey. Do NOT sample from scats this old (Freshness scale D/E)

Old scats look furry if they contain hair, whereas fresh hair-filled scats are still smoothed down by the mucus layer. Old scats without much hair look powdery and white/grey. Do NOT sample from scats this old (Freshness scale D/E)