SHARP SOP 16 Apr 2019 (ECP, past - ARK)

Revised May 21 2019 by Alex Cook, (Original Document-Alison Kocek 23 April 2018)

PASSIVE INTEGRATED TRANSPONDER (PIT) TAG APPLICATION & FIELD RFID USE PROTOCOL

Saltmarsh Habitat & Avian Research Program

Standard Operating Procedure

Survey Goal:

To identify nest ownership and attentiveness of saltmarsh, nelson’s, and seaside sparrows with less disturbance than the use of target netting.

General Approach:

PIT tags are prepared by attaching them to color bands using epoxy. In the field, they are placed on the bird’s leg along with another color band, with the two color bands taped together for support. Later, when a nest is found, the RFID reader is left at the nest for 15-30 minutes to try and get a read from the female’s PIT tag. It allows for knowledge of nest ownership without disturbing the nest by setting up nets every time a new nest is found.

Version history

Date Revised

Summary of Changes

Reason for Change

Author

Notes

2014

Original Document

Alison Kocek

23 April 2018

Updated info about:

RFID reader use

Pictures of Banding Scheme

More information

Alison Kocek

21 May 2019

Removed info about soaking bands and tags in Alcohol

Clarified some language

Added photo of reader on nest

Alcohol soaking wasn’t necessary

Alex Cook

Could use more/better photo of reader antenna on nest

Equipment Needed:

· Biomarker HPT 12 tags (12.5mm x 3mm, 0.115g) $3 each:

· www.biomark.com/products/tags/bulk_pit_tags/

· *RFID Reader and antennae (many options)

· *Hand-held RFID Reader (not required)

· Darvic Color Bands & stirring stick

· Vinyl Electrical Tape colored to match your darvic band color: http://www.3m.com/product/information/Scotch-Super-33plus-Vinyl-Electrical-Tape.html

· Gorilla Glue Epoxy: http://www.gorillatough.com/index.php?page=epoxy

· Nail File

· Scissors

· Toothpick

Frequent recaptures of birds at the nest (as tidal marsh sparrow species may renest several times throughout the season) can lead to nest abandonment and trap avoidance. To reduce both of these costs, attaching a PIT tag to color bands is a novel way to allow researchers to identify adults attending a nest passively. These tags are not powered by a battery and should last the lifespan of the bird. They can also be integrated into future studies looking at nest attendance rates during incubation, feeding rates to young, attendance rates during storms/floods, etc.

Attachment of PIT Tags to Bands:

Attachment of PIT Tags to bands should take place at least 24-hours prior to use in the field.

1. Take a darvic color band (the same color you would normally use) and using your nail file, roughen a thin section of the band located directly opposite the side with the opening (Figure 1a below).

2. Pre slice small strips of of electric tape about the width of a color band to be used to secure PIT tags to bands while epoxy is drying.

3. Open your epoxy glue and mix it per the instructions on the tube. Be sure to mix it for at least 30 seconds, if under mixed the epoxy will not be strong and PIT tags will fall off bands, Let the epoxy sit for ~3-minutes to allow it to become slightly gummy.

4. Using a stirring stick, apply a liberal layer to the section of the band you just roughened, but be sure to not let it drip over the band or into the band (Figure 2).

5. Quickly place the PIT Tag into the glue with the bottom of the PIT Tag sticking out very slightly from the bottom of the band (Figure 1b, 3). Hold for 1-2 minutes. The glue will remain somewhat wet at this point, but should be dry enough that the PIT does not readily fall off the band on its own. Ensure that you can see a small clump of glue on either side of the PIT Tag and that the tag is parallel to the band (not leaning to the side). If needed, more glue can be added to the sides of the PIT Tag to ensure it will be thoroughly attached.

6. Using your pre-cut tape, wrap it tightly around the band, holding the PIT Tag securely onto the band (Figure 1c, 4 ). Place the center of the tape on one side of the band, fold over PIT tag and pinch tape together, being sure not to move the location of the PIT tag. Confirm it is still centered on band.

7. Leave to dry for 24 hours and then remove the tape. Ensure the PIT is securely attached to the band. If it is not, there may have been inconsistency with mixing equal parts of the epoxy glue together. Pull glue off and try again.

Figure 1. At base: (a) Roughen a small part of the band opposite the opening with a nail file. (b) Attach a PIT Tag to the band at the roughened spot using epoxy glue. (c) Once fairly hardened, use a piece of electrical tape to hold the band and PIT together for 24 hours. In the field: (d) Place a darvic band on the bird’s leg and then attach the darvic/PIT band above it. (e) Cover both bands and the PIT in colored electrical tape.

Figure 2. Cover the roughened area on the band with epoxy glue (~3-min after mixing).

Figure 3. Place the PIT Tag into the glue. Align the start of the copper in the PIT tag with the edge of the band.

Figure 4. After ~2-minutes, surround the PIT and band with electrical tape to “clamp” the two together for 24-hours.

Figure 6. Place the PIT/band on the bird with the exposed PIT facing away from the bird’s body

Figure 7. The second band should be placed on the bird to

fill the gap between the PIT and the leg.

Figure 8. Cover both bands/PIT with a single strip of electrical tape. It should be flush with both edges of the band but not extend beyond the edges.

Figure 9. Banding scheme for PIT Tagged birds.

Attachment to Birds in the Field:

1. For our current research objectives, female sparrows of all species and male seaside sparrows will be banded with a PIT Tag as the goal of this study is to reduce the need for target trapping birds on nests. For details on how to band birds, please refer to the SHARP Banding SOP (http://www.tidalmarshbirds.org/wp-content/uploads/downloads/2012/06/SHARP_Banding_SOP.pdf) and the USGS Banding Lab website (http://www.pwrc.usgs.gov/BBL/).

2. Sex the bird to determine if it is a female sparrow (or male seaside sparrow).

3. Check the bird to determine if it is already banded. Banded birds will receive their PIT Tag bands on the opposite leg of their metal band to prevent a metal band damaging a glass PIT Tag. If a darvic and metal band are found on the same leg, the darvic may remain and the PIT-band set-up will be placed on the opposite leg.

4. Newly banded birds will be banded using the banding scheme in Figure 9 ( A = aluminum metal band)*** :

· New female combination: A:PIT-band

· Female previously banded as adult: A/darvic:PIT-band

· Female previously banded as a juvenile or non-breeding adult: PIT-band:A

· New male SESP combination: PIT-band:A

· Male SESP previously banded as an adult: PIT-band:A/darvic

5. Write down the metal band number and PIT Tag RFID # (collected using an RFID Reader which should be present at the banding station with you) on the banding data sheet.

6. Place the metal band on the appropriate leg of the bird if not already banded (see also SHARP banding SOP.

7. Place the PIT-band on the bird (the leg opposite the metal band). The section of the PIT Tag that is not attached to the band should be facing down toward the bird’s foot (Figure 6). If the bird already has a darvic band on the leg which the PIT Tag will be placed, be sure that it the PIT-band is placed above it so that the band in place covers the gap between the PIT and the foot (Figure 7).

8. Place a second darvic band below the PIT-band in order to fill in the gap between the bird’s foot and the PIT Tag (Figure 1d, 7). Ignore this step if a bird already had a darvic band at capture on the PIT-band leg.

9. Cut a long pre-measured piece of electrical tape (8mm in length) and wrap it around the bands and PIT Tag ~3 times while ensuring the bands are pushed together so that no gap is between them. The tape should be about the width of the two bands (a little less than half the width of the electrical tape), covering them both but not extending past them in any way (Figure 1e,8). Pull the tape taut as you wrap it to ensure that there are no gaps or air bubbles and smoosh the tape with your fingers and rub it into the area just around the PIT itself to aid with keeping the tape in place long term. During this process, do your best to touch the sticky part of the tape as little as possible as this will reduce its effectiveness over time.

10. Once a bander is experienced with this technique, this process should take ~1 minute, especially if tape is pre-cut and ready to go when banding.

11. Always double check that the RFID # was written down and confirm the number before releasing a bird. Consider double-checking with the PIT tag reader as well.

*** Note, each master permit holder must get authorization to change the banding status of a bird after its original capture (e.g., adding a color band to a previously banded bird, removing a color band, or adding a PIT Tag band). This may only occur on birds banded by the SHARP project. Aluminum bands should NEVER be moved/removed/replaced. The final banding combination (e.g., the combination the bird will have upon release) will be used on the banding sheet and the original combination of the bird during initial capture will be recorded in the notes. When banding status is changed, there are specific reporting requirements. Using the recapture function, “How Obtained” must be recorded as “Captured by bander, status of bird changed” and “Remarks” should include “Status changed by bander” and a description of the markers that were added. A bird with a color PIT Band will now be status: 325. A status of 325 signifies a healthy bird with a METAL, AUX Marker (color band) and a PIT Tag For more information, see: http://www.pwrc.usgs.gov/bbl/MANUAL/aarespus.cfm and http://www.pwrc.usgs.gov/bbl/manual/statchg.cfm

Use of RFID Reader in the Field (instructions specific to Biomark HPR Plus reader which is recommended):

RFID reader antenna must be placed within ~4 inches of the nest in order to detect an attending adult.

1. Turn on the reader and press the read button to activate it to read tags – it should say “Reading” in green at the top of the screen.

2. Nestle the antennae into the vegetation with the antennae leaning against the nest without pushing it over. (Pictured below)

NOTE: It is recommended to cover antenna and cord with vegetation to conceal its presence.

Antenna can be placed on its side, parallel to nest as well.

3. Use a test PIT Tag such as the fish keychain provided by Biomark (Figure 11) to test if the antenna is close enough to the nest. Do this by gently inserting the test tag into the nest cavity – do not drop it or lay it down on the nest contents. Then check the reader to see if the identity of the test tag was read. The Test Tag ID should show up on the front screen on the reader. Write down this ID so that you don’t confuse it with an actual bird ID in the future. If it does not, reposition the antenna and try again.

4. Once the antenna is properly placed, cover the Reader with a dark colored plastic bag or water-resistant fabric carrying case to keep it safe from water and make it more discrete. Cover the reader and wire leading to the antenna and the antenna itself with loose thatch or wrack collected away from the nest. This is not critical but may help reduce disturbance to returning adults that may be wary of unfamiliar objects near the nest.

5. Record the exact time when you leave the nest site on the Capture Locations Datasheet (http://www.tidalmarshbirds.org/wp-content/uploads/2014/08/2017_SHARP_Capture_Locations-1.pdf). Set a timer for 15 minutes (time can be extended if necessary) and ensure all team members are far away from the nest site so as not to disturb an attending adult from returning to the nest.

6. After 15 minutes have passed, return to the nest site and check to see if there is a new number displayed on the reader. This would be seen as a list of numbers on the front screen (be sure it differs from the test tag ID which was collected when setting up the reader).

7. If there is a read, carefully remove the antenna, reader and all wrack/thatch that was placed by the nest. Once safely away from the nest, you can scroll through the settings to see when the bird first returned to the nest and how long it sat on the nest (see section on downloading data below). Be sure to write this in the notes of the Capture Locations sheet and ADD ALL RFID CAPTURES to your Banding Sheet. Capture time can be found by scrolling through reads on RFID reader which contain time stamps. Be sure you scan through the capture log to see if multiple individual IDs are captured – this can occur with seaside sparrows as males sometimes enter the nest area to feed young or check on the female following a disturbance. This can be done later when out of the field (see Collecting Data section below).

8. If no PIT ID was captured by the reader and you saw a bird flush from the nest and/or you were sure all team members were far enough away from the nest that the bird was not disturbed from returning, the attending adult likely does not have a PIT Tag and will need to be target netted.

9. Once a seaside sparrow nest hatches, the identity of male seaside sparrows may be obtained. Repeat the process above but leave the reader on for 30 minute intervals. Chances of capturing the male ID increase with the number and age of chicks. In NY in 2014, we had a high percentage of our male seaside sparrows banded with PIT Tags and were able to identify males on 86.7% of the nests that reached the chick stage.

Figure 10. Test PIT Tag with PIT Tag ID circled in yellow.

Collecting Data off of the RFID Reader (Biomark HPR Plus):

1. The RFID of captured PIT Tags should show up on the main screen as they are scanned. It is important that the time in and out for all nest collection is recorded so that the RFIDs captured can be appropriately sorted to the correct targeting attempt later. The reader will also maintain records of RFID use at the banding station which can possibly be helpful if an ID was not properly written down on the banding sheet, it can be double checked in the reader’s log.

2. It is beneficial to download data and change your file number daily so to help keep data secure and accurate but this is not essential as all readings will be date and time stamped.

3. Viewing data already collected: Hold down ”OK” to turn on device. Press Menu -> Files. Ignore the top set of “log” files and scroll down to the “txt” files. Click “OK” when the file for the day you want to view is highlighted. Click on Actions and then click the up (^) arrow to View Tags. You can now scroll through all of the tags collected in this file and double check that everything is written correctly in your Capture Locations sheet and transferred properly to your Banding Sheet (don’t forget this important step!).

4. Downloading Data: Download Biomark application from their website. Attach the reader to your computer using the provided cord that goes into the top right knob on the reader. Once connected, press Menu -> Files. Ignore the top set of “log” files and scroll down to the “txt” files. Click “OK” when the file you want to download. Press the button for Actions then press the left arrow to “Send to PC.” The file will download. Repeat on other files as necessary.

5. If you do not have the ability to download files regularly or forget to. You can change your file name from day to do day to keep record keeping easier. To do this: Menu -> Settings->Memory. Scroll down to the File Number, click “OK” and scroll up or down to change the File number to a new number.

6. Other things to know:

· To turn off the reader, hold down “OK” for 5 seconds.

· If your reader glitches and stops working, hold down “OK” for 30+ seconds and this should restart it for you.

· Remember to charge your reader every night as you will probably only get 6-8 hours of continual use a day from a full battery.

· The reader is waterproof but the ports aren’t so make sure the knobs are screwed on tight before using it in the field.

· Wipe down the metal parts with an alcohol wipe at the end of each day to prevent corrosion.

· Remove the antenna every day before exiting the field so you don’t accidentally bend the prongs connecting the antenna to the reader.

Figure 11. Biomark HPR Plus RFID Reader & antenna.

Figure 12. Close-up of RFID Reader screen and buttons.

Literature Review of Attachment Methods:

Review of RFID Application Methods (Subcutaneous and Band Attachment):

Bonter, David N., and Eli S. Bridge. "Applications of radio frequency identification (RFID) in ornithological research: a review." Journal of Field Ornithology 82, no. 1 (2011): 1-10.000000000000.

Tape on 1 Band:

Weimerskirch, H., O. Chastel, Y. Cherel, J-A. Henden, and T. Tveraa. "Nest attendance and foraging movements of northern fulmars rearing chicks at Bjørnøya Barents Sea." Polar Biology 24, no. 2 (2001): 83-88.

Electrical Tape and 2 Bands:

Bridge, Eli S., and David N. Bonter. "A low‐cost radio frequency identification device for ornithological research." Journal of Field Ornithology 82, no. 1 (2011): 52-59.

Epoxy Glue & Electrical Tape on 1 Metal Band:

Cresswell, Will, S. Holt, J. M. Reid, D. P. Whitfield, and R. J. Mellanby. "Do energetic demands constrain incubation scheduling in a biparental species?."Behavioral Ecology 14, no. 1 (2003): 97-102.

Epoxy Glue on 1 Band:

Boisvert, Michael J., and David F. Sherry. "A system for the automated recording of feeding behavior and body weight." Physiology & behavior 71, no. 1 (2000): 147-151.

Unknown Glue on 1 Band:

Mariette, Mylene M., Emma C. Pariser, Amanda J. Gilby, Michael JL Magrath, Sarah R. Pryke, and Simon C. Griffith. "Using an electronic monitoring system to link offspring provisioning and foraging behavior of a wild passerine." The Auk128, no. 1 (2011): 26-35.

Super Glue on 1 Band:

Schroeder, Julia, Ian R. Cleasby, Shinichi Nakagawa, Nancy Ockendon, and Terry Burke. "No evidence for adverse effects on fitness of fitting passive integrated transponders (PITs) in wild house sparrows Passer domesticus."Journal of Avian Biology 42, no. 3 (2011): 271-275.

Unknown Glue to 2 bands:

Ottosson, Ulf, Johan Bäckman, and Henrik G. Smith. "Nest-attenders in the pied flycatcher (Ficedula hypoleuca) during nestling rearing: a possible case of prospective resource exploration." The Auk 118, no. 4 (2001): 1069-1072.

Cotton Wire and Glue on 2 Bands:

Freitag, Anne, Alcherio Martinoli, and Joseba Urzelai. "Monitoring the feeding activity of nesting birds with an autonomous system: case study of the endangered Wryneck Jynx torquilla." Bird Study 48, no. 1 (2001): 102-109.

Suggested Heat Shrink (Used Electrical Tape):

Bridge, Eli S., and David N. Bonter. "A low‐cost radio frequency identification device for ornithological research." Journal of Field Ornithology 82, no. 1 (2011): 52-59.

Heat Shrink Tubing on 2 bands:

Zangmeister, Jennifer L., Mark F. Haussmann, Jack Cerchiara, and Robert A. Mauck. "Incubation failure and nest abandonment by Leach's Storm‐Petrels detected using PIT tags and temperature loggers." Journal of Field Ornithology80, no. 4 (2009): 373-379.

Specially made Celluloid Band with PIT Inside:

Wilkin, Teddy A., Lucy E. King, and Ben C. Sheldon. "Habitat quality, nestling diet, and provisioning behaviour in great tits Parus major." Journal of Avian Biology 40, no. 2 (2009): 135-145.

Attachment to 1 Band (Unknown Method – may be specially made PIT Band):

Pariser, Emma, Jeff Graves, Lucy Gilbert, Lee Astheimer, and Simon Griffith. "Differential female investment in response to male attractiveness: a study of zebra finches in the wild." Wild at heart? (1910): 64.

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