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Collecting tissue and other samples

Sampling and samples shipment
By
Feb 4, 2003, 8:47pm

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The following article is specific to a company who is offering services, but the general information is useful for collections in general.

1.- Objectives.

This page is a support tool for clinical veterinarians. Its purpose is to facilitate and optimize sampling and samples shipment for analysis at EXOPOL.

Here you will find our recommendations for basic sampling material, packaging to be used, as well as the most appropriate preservation and transportation methods.

It is important to take into account that the type of sample, shipment conditions and storage are key factors for correctly interpreting test results.

EXOPOL proposes a broad range of tests: microbiological isolation, antibiograms, serology, parasitology and immunocytochemistry, as required in each case.

Based on the samples received and the veterinarian requests, EXOPOL selects the best techniques for ensuring an optimum cost-diagnosis ratio.

Basic sampling Material

2.- Basic sampling Material.

In order to optimize the possibilities of bacterial isolation, whenever possible the sample should be collected before starting antibiotic treatment.

The organs submitted should come from live animals, animals put to sleep by the veterinarian, or animals that have died recently (2-4 hours maximum), followed by immediate refrigeration. They must be animals that showed signs of disease, though ideally they should not be terminally ill.

The selected samples (milk, blood, serum, organs, swabs, etc.) should be submitted with the most appropriate transport material along with the veterinarian report.

Our recommendations concerning the minimum material that should be available in the working vehicle to ensure adequate sampling are the following:

Various types of swabs

A.- Sterile tubes with airtight sealing (Venojet type).

Without anticoagulant, for the shipment of sera, milk, urine and other fluids. A sufficient sample volume is 2-3 ml.

With anticoagulant:

Heparin, for the shipment of blood samples when requesting bacterial culture or lymphoid cell culture (viremia, tuberculosis, etc.).

EDTA or heparin for the evaluation of blood parasites and the shipment of joint or cerebrospinal fluid.

B.- Airtight bottles and containers

These are used for shipping organs, tissues, feces, etc. Glass bottles should be avoided because they may be broken. Pharmacy urine sampling containers may be used, as well as simple Tupperware available in supermarkets, or any other airtight container. It is important to check for airtight sealing, to avoid leakage during transport. In no case should different organs be placed in the same container, since this would favor bacterial contamination.

C.- Syringes y sterile needles

For the extraction of blood, joint fluid, cerebrospinal fluid, stomach contents, etc.

D.- Airtight bags
Swabs are easy to transport and require no refrigeration; simply attach the report and use an envelope.

To transport organs or fetuses if no airtight containers are available. Thin and resistant plastic bags such as those used to freeze foodstuffs are a practical option. It is essential to first drain the organ to eliminate as much fluid as possible, and to tie several knots to optimize air tightness. Never include different organs in the same bag, and remember to adequately identify each.

E.- Swabs with transport medium (Amies, Stuart or any other).

These can be obtained from distributors of veterinary materials or in pharmacies. The transport medium is a gel that prevents the drying of cells and limits sample deterioration. Swabs are very practical, since they allow samples to be obtained from live animals (rectal, nasopharyngeal, vaginal, ocular, cutaneous swabs, etc.) or from organs after necropsy. Transport is less costly, no refrigeration is required and biological risks are avoided. For correct sampling, the swab should be rotated while rubbing against the tissue walls, to free and trap cells from the surface.

F.- Gloves

These can be used to transport feces by simply turning the gloves inside out after sampling directly from the rectum.

A perfect refrigerated shipment
G.- White cork boxes

Whenever samples are to be shipped under refrigerated conditions, and especially in summer, the use of these boxes is recommended. They are not completely airtight, however, so it is important to make sure that the contents are properly packed.

H.- Permanent marker

To identify containers, plastic bags, tubes or swabs.

I.- Ice-gel blocks

These are essential for the shipment of refrigerated samples. Never use frozen water.

J.- Samples submission sheet

Although this is one of the most important elements of samples shipment, it is often overlooked — a fact that creates serious problems upon deciding which tests to perform in the laboratory. The basic information that must be provided is:

  • Basic material for necropsies
  • Name of the veterinarian and/or company
  • Name of the farm owner
  • Identification of the organs submitted or of the place from which the swab was taken
  • Brief anamnesis of the case, indicating symptoms, treatments and vaccinations
  • Test required or suspected to be necessary

Gloves, scalpel, tweezers, scissors, disinfectant, face mask and a disposable coat are the customary materials required to perform a necropsy

 

K.- Necropsy material

Gloves, scalpel, tweezers, scissors, disinfectant, face mask and a disposable coat are the customary materials required to perform a necropsy.

3.- Sampling

In order to determine which samples to send, you can download the "Practical Sampling Guide" in *.pdf format

Choose animals with clinical symptoms and (where possible) not subjected to treatment.

  • Maximum asepsis should be observed while working.
  • Send the sample to the laboratory as soon as possible.
  • Live animals
  • Use a resistant box.
  • Make several ventilation holes in the box — especially in summer.
  • Do not include either food or water.
  • Place absorbent material at the bottom to collect excrements.
  • Send in the late afternoon.
Blood
  • Whole blood with anticoagulant.
  • Refrigerated at 4ºC; the shipment should reach the laboratory in less than 24 hours after extraction. Protect tubes from any sort of physical damage.
  • For bacterial cultures: EDTA or heparin.
  • For lymphocyte culture (viremia, tuberculosis): only heparin.
  • For blood parasites: any anticoagulant or directly extending a fine blood smear onto a glass slide.
Serum

Requires no refrigeration. Never freeze the sera with blood clot. The clean serum can be frozen after removing the clot. Protect the tubes from physical damage.

In tubes without anticoagulant. To maximize the volume of serum, incline or invert the tube at room temperature for about 30 minutes.

Feces

For coprological analysis

  • Refrigeration not required. Do not freeze. Use an airtight container
  • Sample directly from the rectum with a glove or plastic bag.

For bacteriological cultures

  • Refrigerated, not frozen, and ensuring a minimum transport time. Airtight container
  • Same procedure as above.
Skin shavings
  • For the diagnosis of ectoparasites, bacterial or fungal infections.
  • Refrigeration not required. To be shipped in sterile bottles.
  • Clean and disinfect the skin. Use a sterile scalpel blade to gently rasp the borders of the affected area, collecting hair and skin and deepening until bleeding occurs.
  • Cerebrospinal or synovial fluid
  • Refrigerated and in tubes containing anticoagulant.
  • Sample with a sterile syringe. Ensure optimum asepsis by disinfecting the injection site and avoiding contamination with blood.
Urine
  • In a sterile airtight container; refrigerate but never freeze.
  • Collect under maximally sterile conditions.
Milk
  • Refrigeration not required provided sampling has been performed correctly. Shipment should be made in a sterile narrow-lipped tube (5 mm).
  • Clean the teat as before milking, and discard the first milk emitted. Fill the tube or sterile container.
  • Protect the tubes from physical damage.
Swabs or sampling brushes
  • Refrigeration not required. The swabs should carry transport medium (Amies or Stuart). Immediate shipment to the laboratory is advised, though if they must be stored over the weekend, don’t forget to refrigerate them.
  • Swabs are useful in application to live animals and in necropsies. They allow the sampling of exudates with high cell contents from cavities: nasal, tracheobronchial, endocervical, rectal and conjunctival. Samples can also be obtained from organs after necropsy. The swab should be introduced deeply into the cavity, rubbing against the walls while rotating the swab to collect the required specimen.
  • Avoid the dragging of feces by first emptying the rectum.
  • Avoid the dragging of vaginal mucus by protecting the swab with the speculum until the cervix is reached.
Organs obtained from necropsy
  • Refrigeration required. Do not freeze if bacteriological testing is requested. Freezing should be reserved for cases where IPX immunocytochemistry is contemplated.
  • Perform the necropsy immediately after death, and no later than two hours postmortem.
  • Maximum asepsis should be observed.
  • Ship affected organs either whole or large pieces — each one in a separate container or bag. Bowels should be ligated at the extremes to avoid loss of contents and contamination.
  • The samples must be placed in an airtight container or bag.
Carcasses
  • Refrigeration required. Do not freeze if bacteriological testing is requested. Freezing is only allowed if immunoperoxidase testing is required.
  • Keep in mind that contamination is likely if the dead tissues have been left more than four hours without refrigeration. Be sure to place samples in an airtight container or bag.
  • The shipping of whole carcasses should be avoided whenever possible— especially in summer. Organs may be sent separately (except in the case of fetuses of small size).

4.- Recommendations for shipment.

The samples must ALWAYS be accompanied by:

  • Veterinarian data.
  • Identification of the farm.
  • Identification of all organs submitted, indicating the species. In the case of swabs, indicate where the sample was obtained.
  • Brief anamnesis.
  • Test requested or suspected clinical diagnosis.
  • Should refrigeration be required, use gel or dry ice blocks instead of frozen water.
  • http://www.exopol.com/in/toma.in.html

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