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Tortoise Faecal Sample Examination

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Tortoise Faecal Sample Examination....

Carrying out your own faecal sampling is not difficult and can be a very useful skill to have to assist in the care for your tortoises. There is an initial investment to make for the required equipment, but you’ll find that this will pay off in no time as you will be able to check your tortoises for worms quicker than if you were to take it to the vets. You will also save on not paying your vet for faecal sampling.

Health and Safety:

Faeces may contain hazardous pathogens (bacteria, viruses etc). Appropriate hygiene and safety procedures should be employed. Local health and safety regulations should be observed.

Collection Of Faecal Samples

Samples should be collected and sampled as soon as possible or dispatched to a laboratory in suitable containers such as:

Screw cap bottles
Plastic containers with lids
Sealed plastic bags

Each sample should be clearly labelled with your tortoises name and date of collection. If your sample is not going to be examined straight away, it should be immediately stored in the refrigerator (4 °C) until it is processed. SAMPLES SHOULD NEVER BE KEPT IN THE FREEZER.

There are a number of different methods available for separating, concentrating and demonstrating eggs in faecal samples. The method described below is the one we use:

Simple Test Tube Floatation

Simple Test Tube Floatation

The simple test tube flotation technique is a qualitative test for the detection of nematode and cestode eggs. This is a useful method to use in preliminary surveys to establish which parasite groups are present.

Equipment List

Two beakers or plastic containers
Disposable Gloves
Tea strainer or double layer cheesecloth
2 or 3 Measuring cylinders or container graded by volume
Fork, craft sticks or stirring rod
Test tube, 16 x 150mm (that holds 20ml in volume)
Test tube holder
Microscope with 4x, 10x & 40x power with a built in light
Blank microscope slides
Coverslips
Scales (that measures in grams)
Timer
Floatation solution (see instructions below on how to make this solution).

Weigh or measure using pre-calibrated scales, 3g (approx ¾ tsp) of faeces and put into a small cup or container that can hold 30-50ml.

Mash the faeces really well with a craft stick or fork. Then slowly pour in 25 ml of flotation fluid.

Stir the faeces and flotation fluid mix thoroughly with a craft stick or fork, then leave to sit for 2 minutes.

Pour the faecal suspension through a tea strainer or double layer of cheesecloth into a larger container ensuring that you force as much of the solution through as possible, then stir.

Let sit for another 2 minutes.

Pour the faecal suspension into a test tube supported in a stand or rack from the container.

Fill it a little more than full so that the test tube is gently topped off with the suspension leaving a convex at the top of the tube (don’t worry if it slightly overflows as it should kind of stand up higher than the top of the test tube)

Carefully place a coverslip on the top of the test tube. The solution should touch the coverslip.

Leave to stand for 15-20 minutes. The eggs will float to the top of the solution and collect on the coverslip.

Carefully lift the coverslip off the test tube together with the drop of fluid adhering to it.

Place the coverslip (wet side down) onto a clean slide.

Place the slide onto your microscope and examine using a 10 x 10 magnification.

Find a corner of the coverslip to start your examination at. Start searching the slide using a upward and downward pattern technique like the one shown below.

You can start looking for worm eggs. Make sure every time you see a worm egg, you make a note on a bit a paper.

The eggs are very small so be patient and keep looking. You will see many things in the sample that you think looks like worm eggs but aren’t. Just keep looking because you will eventually spot one. You are looking for oval shaped eggs. If you think you have come across a worm egg, you can then switch your magnification on your microscope up to 40X to have a closer look.

Once you have finished your examination, count the total number of times you made a note on your bit of paper. This will be the total amount of eggs you saw.

How To Make Your Own Floatation Fluid

You are about to make a saturated solution. This entails mixing as much Epson Salt into some water until the water cannot take any more. This will cause the water/salt solution to become heavier than standard water hence how the worm eggs will float to the top of the solution when you carry out your examination.

Purchase some Epson Salt.
Pour some Epson Salt into a jar that has a tight fitting lid then fill about 2/3 full of water.
Screw the lid on tight. Shake the jar well.
Add some more Epson Salt and shake well.
Over a 24 hour period, keeping adding more Epson Salt and shaking the jar until you have a permanent layer in the bottom of your jar. Once you have this permanent layer, it means no more salt will dissolve in the water.
You now have a Epson Salt floatation solution.
You now have to pour off the solution into a fresh jar, leaving behind the un-dissolved Epson Salt.
You now have a floatation solution ready for use in your examination.

Pictured below is a very good book to use as reference when carrying out your faecal sample examinations. The book is titled 'Understanding Reptile Parasites' 2nd Edition by Roger J. Klingenberg. The book certainly helped us to learn all about the different types of parasites that can be found and what they looked like.

Understanding Reptile Parasites (Herpetocultural Library)

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