Wing Punch and Hair Sampling Protocols
Tissue and hair samples can be taken from live bats. Follow normal protocols for safe and humane handling of the animals. If you are going to take wing punches or hair samples, plan ahead and make sure you have the necessary equipment.
List of Equipment:
- Lighter (to flame instruments)
- Vials containing storage solution for membrane punches
- Empty vials for hair samples
- Storage box for vials
- Fine-point or tissue forceps
- Iris scissors
- Biopsy punches (3 mm)
- Bottle of alcohol or alcohol swabs for wiping instruments and surface
- Latex gloves (optional)
- To request vials for storing samples, contact Dr. Nancy Simmons (firstname.lastname@example.org).
- Biopsy punches can be obtained from many sources. One supplier is VWR.
Wing punches are small (3mm) circles of skin removed from the wing membrane using a biopsy punch. Based on recaptures of sampled bats, the holes in the membrane usually grows back within 2-3 weeks, so there are no long-term effects. Bats are commonly captured while mistnetting with holes in their wings that are much larger than those inflicted by wing punching, and these holes don’t appear to result in a loss of flight ability. When taking tissue from the wing membranes, take the samples from close to the body (between the leg and the fifth digit in the wing); this is thought to minimize the effect on flight performance. Do not punch areas with large blood vessels.
- Flame the biopsy punch and forcep thoroughly to sterilize them and ensure that no tissue or hair from the last bat remains. The instruments should get hot.
- Let the instruments cool by placing them on the vial box in such a way that the business ends do not touch anything and therefore remain sterile. If you don’t let them cool, you will cauterize the bat’s skin when you take the punch, which may prevent proper healing of the hole.
- Wipe the instruments with an alcohol swab to remove any residue from the flaming and let the instruments dry for a few seconds.
- Remove the bat from its holding bag and stretch the wing over a flat, hard or semi-hard surface (cutting board, clipboard, binder, cardboard, etc.). While the membrane is stretched, press the punch down onto the membrane of one wing close to the legs (between the legs and the fifth digit), and twist and/or rock the punch slightly until you can tell the punch has gone through the membrane on all sides. There is no need to hammer the punch down through the membrane, and doing so will decrease the life of the punch. Each punch can be reused multiple times (5-40 depending on how hard you are on it), but please use your judgement as to how well the punch is cutting, and dispose of punches as soon as they start to dull.
- The cut tissue will now be sitting on the surface you punched on, or may be in the hollow portion of the punch. If the wing tissue is still in the punch, use the forceps to extract it. Transfer the membrane to an O-ring vial containing liquid preservative. The tissue tends to stick to the forceps, so you might have to shake the forceps semi-vigorously in the solution in the vial to dislodge the sample, or wipe it off onto the side of the vial.
- Repeat for the other wing. Place both pieces of membrane from an individual into the same. When finished, please make sure that both pieces of tissue are sitting in the solution. You may have to shake the vial (with the cap on!) to dislodge them from the sides of the vial.
- Make sure to label all vials with your unique identifier for that bat, the date (day/month/year, with the month written out, e.g., 12/Aug/2001, or Aug/12/2001), bat species, sex, reproductive condition, and age. Please also fill out the data sheet provided with the necessary information. Please do not write on the cap.
- Between bats, please make sure that you clean the punching surface well, either by flushing with a spray bottle containing alcohol (70-95% ethanol or isopropyl) or wiping down the surface well with an alcohol swab. The goal is to minimize the chances of contaminating future samples.
- If you ever have the opportunity to collect from dead bats, please collect a decent amount of membrane from each wing (1cm x 1 cm area) and place it in a vial with preservative. Please also take some muscle tissue (it is easiest to take it from the pectoral muscles) and store it in a separate vial with preservative. Take a minimum of a 2 mm3 piece of tissue (a small cube), but if you can, collect as much as will fit into the vial and still allow sufficient solution to preserve the specimen. Do not overstuff vials; use multiple vials for the same individual if necessary.
- Clean the scissors by dipping in alchol or wiping them with an alchol swab. If you are in doubt as to their cleanliness, flame the scissors as described above under the wing punch protocol. Allow them to cool and dry.
- Clip a small amount of fur (1 cm x 1 cm area) from the area between the scapulae using scissors. Get as much of the length of the hair as possible, but you do not necessarily have to cut down to the base.
- Store the hair in an empty vial. Do not put hair into liquid preservative.
- Label the vial with your unique identifier for that bat, the date (day/month/year, with the month written out, e.g., 12/Aug/2001, or Aug/12/2001), bat species, sex, reproductive condition, and age. Please also fill out the data sheet provided with the necessary information. Please do not write on the cap.
- Once finished, please wipe any remaining hair off of the scissors with an alcohol swab. Be very careful to avoid cross-contamination.