FEATHER SAMPLING PROTOCOL

Transcription

1 American Kestrel Genoscape Project University of California Los Angeles, Boise State University, HawkWatch International, the American Kestrel Partnership FEATHER SAMPLING PROTOCOL Our plan - We have completed genomic sequencing, and are developing primers for assays of hundreds of samples. We will be building population specific migration maps within the next couple of years for American kestrels. Please help us collect samples from American kestrels across their range and annual cycle. Our request - We welcome feathers from migratory or resident kestrels, collected any time of the year, including breeding, migration, or winter. When body feathers are pulled, a small amount of skin cells remain attached to the quill of the feather. These skin cells are a valuable source of DNA that can be used to determine the population origin of an individual bird. We recommend that two body feathers be collected during the banding process from each bird. There is no need to collect feathers from the same individual more than once. And there is no need to collect samples from every nestling in a brood. One nestling will suffice. Necessary equipment and required permits To collect feathers, you will need labeled sampling envelopes (which we will send to you when you sign up) and a fine tip pen. In the US, to collect 4 body feathers from American kestrels, banders will need a federal bird banding permit from the USGS Bird Banding Laboratory (BBL) that gives permission to take, possess, and transport feather samples (for how to get permission from the USGS BBL please see below). In addition, you will likely need permission from the State agency where you are collecting feathers. For more information about State specific requirements please visit: You do not need a USFWS scientific collecting permit to collect body feathers from live birds while you are banding, if you have permission from the Bird Banding Lab ( In Canada, to collect 4 body feathers from American kestrels: 1) If you already have a Canadian Bird Banding Permit from the Canadian Bird Banding Office (Environment and Climate Change Canada) you will need to add proper authorization to take, possess and transport feather samples AND you will need a Scientific Collection permit from your province/territory 2) If you do not have a Canadian Bird Banding Permit, you can apply for a Scientific Collection/Wildlife Research permit from your province/territory to collect feather samples from American Kestrels

2 Countries outside of the United States and Canada will have different permit requirements to collect American Kestrel feathers depending on laws and regulations in that country. Please contact your local wildlife agency to determine requirements. If you are unsure whether you meet the above requirements, and would like help navigating the federal, state or international permitting systems, we can help! Please Contact Us ***Copies of federal bird banding permits and state permits must be included in sample shipment*** Collecting Feather Samples From live birds: To collect a sample, gently tease apart feathers on the belly/breast of the bird until you have a feather firmly between your thumb and finger as close to the body as possible. To pluck the feather, just hold them relatively close to the base, and pull quickly and gently. Ensure that you are pulling contour feathers, rather than down feathers, as these provide quality DNA (see pictures and diagram below). Do not touch the quill, as the DNA is extracted from the skin cells attached to it. Place the feathers, 4-5 total, from each bird into a labeled sampling envelope. Providing your own envelopes is fine, but please make sure each envelope contains the necessary data described below. American kestrel contour feathers (left) and down feathers (right). Please only collect contour feathers, as these provide quality DNA for genetic analyses.

3 Diagram of bird feather types. Please collect contour feathers from the belly or breast, as these provide the best quality DNA for genetic analyses. From carcasses: Feather samples from live birds are preferred, as they provide high quality DNA for analysis. However, feathers can be taken from kestrel carcasses in cases where the kestrel is recently deceased, but the collection method is slightly different. In these cases, please pull out a clump of feathers, ensuring that a visible amount of skin is attached to the tips. Please note that collection permits are still required whether feathers are taken from live or dead birds, and the number of feathers in the clump should be recorded. Feathers can be stored at room temperature. Please send sample shipments at the end of the field season (see instructions below). Required Data 1) On each envelope please provide the following information: Date (Please use letters for the month instead of numbers, e.g. Apr 25, 2016) Species Name Band Number Location (closest town, State/Province, country [eg: Boise, ID, USA]) Sex (male or female) Age code: adult (A; any non-nestling bird capable of flight) or nestling (N; birds found in the nest) [if nestling please record age in days in the UKN line] Please check yes for Breeding If samples are from an adult bird captured on the nest or at a territory, otherwise check No or UNK. Recapture If the bird was previously banded by you or another bander (yes) or the bird is newly banded (no)

4 UCLA # (leave blank) 2) In addition to the data on the sampling envelopes, please fill out the excel data sheet below. Please this data sheet to fcphenology@boisestate.edu and include a hard copy with your shipment. Note: GPS coordinates: Please provide latitude and longitude in decimal degrees based on a WGS84 projection (to 6 decimal places, e.g , ) Example Date 30- Jun- 17 Band Number Latitude Longitud e State/ Provinc e Countr y Age (Adult or Nestling Nestling age (in days) Idaho USA Nestling 23 S e x m a l e Number of feathers Bander s Name 4 Julie Heath

5 Shipping Feather Samples For shipments within the U.S., please include the following: 1. A copy of the importers (Boise State University) USDA permit # for shipments within the US [last 2 pages of this doc] 2. A copy of your (the collectors) federal bird banding permit and state permit (as necessary). 3. A datasheet with the above information 4. The samples For international shipments (i.e. from countries outside of the U.S.) Please contact us before shipping samples to ensure the proper permits and procedures are in place so that samples do not get held up at the border* In addition to the above, you will need to include two importers (Boise State University) USDA permits # and #133611, and a CITES export permit to ship samples, which we will help fill out and obtain before you ship samples. Some countries/states/provinces also require other export permits. Returning Completed Samples to Boise State with necessary paperwork Send your samples in a single shipment at the end of your field season using the provided return FedEx shipping labels (the address is also provided below). If shipping without the provided label please send via FedEx or UPS. PLEASE DO NOT USE THE U.S. postal service to send us your feather samples. The U.S. Postal Service irradiates mail with high-power radiation that might damage the DNA. SHIP TO: Julie Heath, Boise State University, 1910 University Dr., Dept of Biol. Sci. Science Building, Room 217A, Boise, Idaho, , phone: , or ; E- mail: fcphenology@boisestate.edu ***Permits must be included in sample shipment*** Thank you very much for your participation in this important work! Please let us know if you have any questions. For additional information or questions, please contact: Anjolene Hunt or Michaela S. Brinkmeyer Boise State University Tel: (208) ; fcphenology@boisestate.edu website: fullcyclephenology.com/

6 How to get permission to collect, possess, and transport American kestrel feather samples on a USGS BBL bird banding permit ( Tissue Sampling (feathers) Please allow 2 months lead time to process your revision request. To request permission to take feather samples, please provide the BBL the following information. 1. Master's name and permit number, and any subpermits that require this permission 2. Species 3. What feathers will be taken, and how many 4. Project proposal (Why are the feathers needed? How will they be used?) This information can be sent to bbl_permits@usgs.gov. Please feel free to use the following text for the permit applications. Research Project Summary Using high-resolution genetic markers to reveal American Kestrel connectivity: towards understanding kestrel population declines and the impacts of climate change on raptor monitoring and management Summary American Kestrels (Falco sparverius) are declining for unknown reasons and studies regarding their rates of survival and migratory ecology are lacking. Further, patterns of bird migrations are changing both spatially and temporally because of climate change. Historically, the challenge for studying migrant ecology, the drivers of regional population declines, and the impacts of climate change has been an inability to link breeding, wintering, and migrant populations. Recently, Ruegg et al (2014) described a new single nucleotide polymorphism (SNP) based method for identifying populations of migratory birds at finer spatial scales than has previously been possible. By harnessing the power of this new SNP-based methodology we will finally be able to tackle long-standing questions in migrant ecology and population dynamics of the American Kestrel. Understanding the migratory connectivity of populations of American Kestrels, and how connectivity changes with climate, will allow us to: 1) identify the wintering grounds and migratory routes of declining breeding populations,2) test hypotheses about population change and 3) assess the impacts of climate change on the effectiveness of migration counts as a tool for monitoring raptor populations. To do this, we will need to collect DNA samples using plucked feathers from American kestrels across their range and throughout the annual cycle. Two to five body feathers will be gently plucked from the belly or back of the kestrel, stored in an envelope, and shipped to UCLA for analysis.

7 United States Department of Agriculture Animal and Plant Health Inspection Service Veterinary Services National Center for Import and Export Animal Products 4700 River Road Unit 40 Riverdale, MD Telephone: (301) FAX: (301) Julie Heath / Boise State University 1910 University Dr., Dept of Biol. Sci. Science Building, Room 217A Boise, ID Friday, April 21, 2017 Dear Julie Heath: Your USDA Veterinary Permit to import and/or transport controlled materials, organisms, or vectors accompanies this cover letter. Review this permit carefully, as the statements and language may have changed to reflect the requirements of newly published regulations. Please note the following: USDA Veterinary Permits no longer require a signature. Use of the permit for importation of the described commodity(ies) is acknowledgement that the permittee is legally responsible for complying with the permit conditions. Review the import permit for errors. Should you identify any errors, please contact our office immediately A copy of the permit must accompany every shipment. Do Not send the permit back to this office. Contact our office with any questions or concerns at , option 1. Sincerely, Dr. Linda Kahn-Tobin Senior Staff Veterinarian Import Animal Products National Import Export Services (NIES) *USER FEES: New permit application $150.00, Renewal permit $97.00, Amended permit $75.00, FBS inspection $ (all fees are per application) and the Import Compliance fee $ per shipment. Safeguarding Animal Health APHIS is an agency of USDA's Marketing and Regulatory Programs An Equal Opportunity Provider and Employer Federal Relay Service (Voice/TTY/ASCII/Spanish)

8 U.S.DEPARTMENT OF AGRICULTURE ANIMAL AND PLANT HEALTH INSPECTION SERVICE VETERINARY SERVICES RIVERDALE, MARYLAND file:///d:/inetpub/wwwroot/epermits/images/ UNITED STATES VETERINARY PERMIT FOR IMPORTATION AND TRANSPORTATION OF CONTROLLED MATERIALS AND ORGANISMS AND VECTORS NAME AND ADDRESS OF SHIPPER(S) Various shippers outside the UNITED STATES PERMIT NUMBER Research DATE ISSUED DATE EXPIRES 04/19/ /19/2018 CC: Service Center, ID (Boise, ID) FWS (Arlington, VA) NAME AND ADDRESS OF PERMITTEE INCLUDING ZIP CODE AND TELEPHONE NUMBER Julie Heath Boise State University 1910 University Dr., Dept of Biol. Sci. Science Building, Room 217A Boise, Idaho / U.S. PORT(S) OF ARRIVAL AS APPLICABLE MODE OF TRANSPORTATION ANY AS REQUESTED IN YOUR APPLICATION, YOU ARE AUTHORIZED TO IMPORT OR TRANSPORT THE FOLLOWING MATERIALS Avian blood, claw clippings, and feather samples RESTRICTIONS AND PRECAUTIONS FOR TRANSPORTING AND HANDLING MATERIALS AND ALL DERIVATIVES THIS PERMIT IS ISSUED UNDER AUTHORITY CONTAINED IN 9 CFR CHAPTER 1, PARTS 94,95 AND 122. THE AUTHORIZED MATERIALS OR THEIR DERIVATIVES SHALL BE USED ONLY IN ACCORDANCE WITH THE RESTRICTIONS AND PRECAUTIONS SPECIFIED BELOW (ALTERATIONS OF RESTRICTIONS CAN BE MADE ONLY WHEN AUTHORIZED BY USDA, APHIS, VS). o Adequate safety precautions shall be maintained during shipment and handling to prevent dissemination of disease. o With the use of this permit I, Julie Heath, Permittee, acknowledge that the regulated material(s) will be imported/transported within the United States in accordance with the terms and conditions as are specified in the permit. The Permittee is the legal importer/recipient [as applicable] of regulated article(s) and is responsible for complying with the permit conditions. The Permittee must be at least 18 years of age and have and maintain an address in the United States that is specified on the permit; or if another legal entity, maintain an address or business office in the United States with a designated individual for service of process; and serve as the contact for the purpose of communications associated with the import, transit, or transport of the regulated article(s). **Note: Import/Permit requirements are subject to change at any time during the duration of this permit. continued on subsequent page(s)... TO EXPEDITE CLEARANCES AT THE PORT OF ENTRY, BILL OF LADING, AIRBILL OR OTHER DOCUMENTS ACCOMPANYING THE SHIPMENT SHALL BEAR THE PERMIT NUMBER SIGNATURE Linda Kahn-Tobin TITLE NO. LABELS National Import Export Services VS FORM 16-6A (MAR 95) Replaces VS Form 16-3A and which are obsolete Page 1 of 3

9 U.S.DEPARTMENT OF AGRICULTURE APHIS / VETERINARY SERVICES, RIVERDALE, MARYLAND ATTACH TO U.S. VETERINARY PERMIT RESTRICTIONS AND PRECAUTIONS: (continued from Permit Form VS 16-6) o ***Each shipment shall be accompanied by an ORIGINAL signed document from the producer/shipper/exporter confirming that the exported material: 1) was derived only from avians (birds), and 2) was subjected to one of the following viral inactivation treatments prior to shipment to the United States: (a) heated to a minimum of 56 C for at least 3 hours, (b) heated to a minimum of 60 C for at least 30 minutes, (c) heated to a minimum of 100 C for at least 20 minutes, (d) treated with a minimum of 2% sodium dodecyl sulfate (SDS), (e) treated with a minimum of 3% beta propiolactone for 12 hours at 4 C at ph 7, (f) immersed in a minimum of 10% formalin, (g) immersed in a minimum of 70% alcohol, (h) immersed in phenol/chloroform mixture, (i) treated with proteinase K, (j) treated with guanidine HCl, OR (k) spotted on Whatman FTA cards (for liquids only). [This certification must CLEARLY correspond to the shipment by means of an invoice number or shipping marks or lot number or other identification method. An English translation must be provided.] o ***Materials shall be consigned directly to the permittee at the permittee address specified above. Materials imported under this permit may be hand carried in personal baggage from the country of origin to the port of arrival, but must be declared and made available to port officials for inspection, and must be transported directly to the permittee by someone with identification and current, signed written authority from the permittee. The permittee's authorizing document must be original, on letterhead, and specific to the particular shipment(s), and shall be valid for no more than 2 months from the date of issuance. o This permit DOES NOT authorize direct or indirect exposure of or inoculation into domestic or laboratory livestock (including but not limited to: birds/poultry/eggs, cattle, sheep, goats, swine, and/or horses). Work shall be limited to in vitro uses only. o Packaging, containers, and all equipment in contact with these materials shall be sterilized or considered a biohazard and be disposed of accordingly. o THIS PERMIT IS VALID ONLY FOR WORK CONDUCTED OR DIRECTED BY YOU OR YOUR DESIGNEE IN YOUR PRESENT U.S. FACILITY OR APPROPRIATELY INSPECTED LABORATORY. THE AUTHORIZED IMPORTED MATERIAL(S) MUST BE SHIPPED/CONSIGNED DIRECTLY TO THE ADDRESS OF THE PERMITTEE AS IDENTIFIED ON THIS PERMIT. (MATERIALS SHALL NOT BE MOVED TO ANOTHER U.S. LOCATION, OR DISTRIBUTED WITHIN THE U.S., WITHOUT USDA, APHIS, VS, NIES AUTHORIZATION.) ++ EXCEPTION ++ Materials that have been treated with one of the viral inactivation methods specified above may be distributed if accompanied by a signed document confirming the viral inactivation treatment that has been applied. Records of distribution and treatments shall remain on file and made available to USDA upon request. o On completion of your work, all permitted materials and all derivatives therefrom shall be destroyed. Page 2 of 3

10 U.S.DEPARTMENT OF AGRICULTURE APHIS / VETERINARY SERVICES, RIVERDALE, MARYLAND ATTACH TO U.S. VETERINARY PERMIT RESTRICTIONS AND PRECAUTIONS: (continued from Permit Form VS 16-6) o Imported material may be subject to regulations enforced by the United States Department of Interior, Fish and Wildlife Service (FWS). Importer must contact FWS, information is available at web pages and/or o Imported articles, such as nests and crop contents, containing plant material may be subject to regulations enforced by USDA, APHIS, Plant Protection and Quarantine (PPQ). Importers of such materials should consult PPQ at: (301) or toll free at: (866) for more information. o This permit does not exempt the permittee from responsibility for compliance with any other applicable federal, state, or local laws and regulations. o The restrictions on this permit remain in force as long as the material is in the United States. o Any person who VIOLATES the terms and conditions of permits, and/or who forge, counterfeit, or deface permits may be subject to criminal and civil penalties in accordance with applicable law. In addition, all current permits may be cancelled and future permit applications denied. o A copy of this permit must be included with the shipping documents. For imported materials, these documents must be presented to CBP Agricultural Specialists upon arrival at the U.S. port of arrival. Page 3 of 3

11 United States Department of Agriculture Animal and Plant Health Inspection Service Veterinary Services National Center for Import and Export Animal Products 4700 River Road Unit 40 Riverdale, MD Telephone: (301) FAX: (301) Julie Heath / Boise State University 1910 University Dr., Dept of Biological S Science Building, Room 217A Boise, ID Friday, April 21, 2017 Dear Julie Heath: Your USDA Veterinary Permit to import and/or transport controlled materials, organisms, or vectors accompanies this cover letter. Review this permit carefully, as the statements and language may have changed to reflect the requirements of newly published regulations. Please note the following: USDA Veterinary Permits no longer require a signature. Use of the permit for importation of the described commodity(ies) is acknowledgement that the permittee is legally responsible for complying with the permit conditions. Review the import permit for errors. Should you identify any errors, please contact our office immediately A copy of the permit must accompany every shipment. Do Not send the permit back to this office. Contact our office with any questions or concerns at , option 1. Sincerely, Dr. Linda Kahn-Tobin Senior Staff Veterinarian Import Animal Products National Import Export Services (NIES) *USER FEES: New permit application $150.00, Renewal permit $97.00, Amended permit $75.00, FBS inspection $ (all fees are per application) and the Import Compliance fee $ per shipment. Safeguarding Animal Health APHIS is an agency of USDA's Marketing and Regulatory Programs An Equal Opportunity Provider and Employer Federal Relay Service (Voice/TTY/ASCII/Spanish)

12 U.S.DEPARTMENT OF AGRICULTURE ANIMAL AND PLANT HEALTH INSPECTION SERVICE VETERINARY SERVICES RIVERDALE, MARYLAND file:///d:/inetpub/wwwroot/epermits/images/ UNITED STATES VETERINARY PERMIT FOR IMPORTATION AND TRANSPORTATION OF CONTROLLED MATERIALS AND ORGANISMS AND VECTORS NAME AND ADDRESS OF SHIPPER(S) Various shippers within the UNITED STATES PERMIT NUMBER Research DATE ISSUED DATE EXPIRES 04/21/ /21/2018 CC: Service Center, ID (Boise, ID) FWS (Arlington, VA) NAME AND ADDRESS OF PERMITTEE INCLUDING ZIP CODE AND TELEPHONE NUMBER Julie Heath Boise State University 1910 University Dr., Dept of Biol. Sci. Science Building, Room 217A Boise, Idaho / U.S. PORT(S) OF ARRIVAL TRANSPORT PERMIT MODE OF TRANSPORTATION ANY AS REQUESTED IN YOUR APPLICATION, YOU ARE AUTHORIZED TO IMPORT OR TRANSPORT THE FOLLOWING MATERIALS Wild avian specimens (previously imported) - treated RESTRICTIONS AND PRECAUTIONS FOR TRANSPORTING AND HANDLING MATERIALS AND ALL DERIVATIVES THIS PERMIT IS ISSUED UNDER AUTHORITY CONTAINED IN 9 CFR CHAPTER 1, PARTS 94,95 AND 122. THE AUTHORIZED MATERIALS OR THEIR DERIVATIVES SHALL BE USED ONLY IN ACCORDANCE WITH THE RESTRICTIONS AND PRECAUTIONS SPECIFIED BELOW (ALTERATIONS OF RESTRICTIONS CAN BE MADE ONLY WHEN AUTHORIZED BY USDA, APHIS, VS). o Adequate safety precautions shall be maintained during shipment and handling to prevent dissemination of disease. o With the use of this permit I, Julie Heath, Permittee, acknowledge that the regulated material(s) will be imported/transported within the United States in accordance with the terms and conditions as are specified in the permit. The Permittee is the legal importer/recipient [as applicable] of regulated article(s) and is responsible for complying with the permit conditions. The Permittee must be at least 18 years of age and have and maintain an address in the United States that is specified on the permit; or if another legal entity, maintain an address or business office in the United States with a designated individual for service of process; and serve as the contact for the purpose of communications associated with the import, transit, or transport of the regulated article(s). **Note: Import/Permit requirements are subject to change at any time during the duration of this permit. o ***Each shipment shall be accompanied by an ORIGINAL signed document from the producer/manufacturer confirming that the transported material: 1) was derived only from avians (birds); and 2) was subjected to one of the following viral inactivation treatments prior to transport: (a) heated to a minimum of 60 degrees C for at least 30 minutes, (b) immersed in a minimum of 10% formalin (c) immersed 70% alcohol,... [continued on page 2]... continued on subsequent page(s)... TO EXPEDITE CLEARANCES AT THE PORT OF ENTRY, BILL OF LADING, AIRBILL OR OTHER DOCUMENTS ACCOMPANYING THE SHIPMENT SHALL BEAR THE PERMIT NUMBER SIGNATURE Linda Kahn-Tobin TITLE NO. LABELS National Import Export Services VS FORM 16-6A (MAR 95) Replaces VS Form 16-3A and which are obsolete Page 1 of 3

13 U.S.DEPARTMENT OF AGRICULTURE APHIS / VETERINARY SERVICES, RIVERDALE, MARYLAND ATTACH TO U.S. VETERINARY PERMIT RESTRICTIONS AND PRECAUTIONS: (continued from Permit Form VS 16-6) o... [continued from page 1]... (d) immersed in phenol/chloroform, (e) heated to a minimum of 100 degrees C for at least 20 minutes, (f) heated to a minimum of 56 degrees C for at least 3 hours, (g) treated with a minimum of 3% beta propiolactone for 12 hours at 4 C at ph 7, (h) immersed in a minimum of 2 percent sodium dodecyl sulfate (SDS) for at least 30 minutes, (i) proteinase K, (j) guanidine HCl and/or (k) spotted on Whatman FTA cards (for liquids only). [This certification must CLEARLY correspond to the shipment by means of an invoice number or shipping marks or lot number or other identification method. An English translation must be provided.] o This permit DOES NOT authorize direct or indirect exposure of or inoculation into domestic or laboratory livestock (including but not limited to: birds/poultry/eggs, cattle, sheep, goats, swine, and/or horses). Work shall be limited to in vitro uses only. o Packaging, containers, and all equipment in contact with these materials shall be sterilized or considered a biohazard and be disposed of accordingly. o THIS PERMIT IS VALID ONLY FOR WORK CONDUCTED OR DIRECTED BY YOU OR YOUR DESIGNEE IN YOUR PRESENT U.S. FACILITY OR APPROPRIATELY INSPECTED LABORATORY. THE AUTHORIZED IMPORTED MATERIAL(S) MUST BE SHIPPED/CONSIGNED DIRECTLY TO THE ADDRESS OF THE PERMITTEE AS IDENTIFIED ON THIS PERMIT. (MATERIALS SHALL NOT BE MOVED TO ANOTHER U.S. LOCATION, OR DISTRIBUTED WITHIN THE U.S., WITHOUT USDA, APHIS, VS, NIES AUTHORIZATION.) ++ EXCEPTION -Material is authorized to be distributed for evaluation, provided the transported material is accompanied by a signed document confirming that the material was subjected to one of the viral inactivation treatments listed above. Records of treatment and distribution shall remain on file and be made available to USDA upon request. o On completion of your work, all permitted materials and all derivatives therefrom shall be destroyed. o Imported material may be subject to regulations enforced by the United States Department of Interior, Fish and Wildlife Service (FWS). Importer must contact FWS, information is available at web pages and/or o This permit does not exempt the permittee from responsibility for compliance with any other applicable federal, state, or local laws and regulations. o The restrictions on this permit remain in force as long as the material is in the United States. Page 2 of 3

14 U.S.DEPARTMENT OF AGRICULTURE APHIS / VETERINARY SERVICES, RIVERDALE, MARYLAND ATTACH TO U.S. VETERINARY PERMIT RESTRICTIONS AND PRECAUTIONS: (continued from Permit Form VS 16-6) o Any person who VIOLATES the terms and conditions of permits, and/or who forge, counterfeit, or deface permits may be subject to criminal and civil penalties in accordance with applicable law. In addition, all current permits may be cancelled and future permit applications denied. o A copy of this permit must be included with the shipping documents. Page 3 of 3

15

16 Text and photograph copyright 2007 All Rights Reserved To inquire about permission to reproduce text or photographs, send an to or Printed and distributed in the United States of America by Zip Publishing, Columbus, Ohio

17 This work is dedicated primarily to all the diligent conservationists and volunteers, the unsung heroes who act behind the scenes, doing the field work for the greater good. 2

18 Synopsis The purpose of this work is to illustrate, through digital photography, important anatomical features, and personal observations, the key developmental stages of American Kestrel nestlings from time hatched until fledged. This work will assist anyone working in the field with these falcons, to provide an easy, accurate method of aging young nestlings, and to better plan for nest box visits and banding. This booklet is meant for anyone with an interest in these falcons: from active conservationists, volunteers and students, to avid and amateur bird watchers or anyone displaying a general interest in or appreciation of birds. 3

19 THE TIMELINE PROJECT Photographic Atlas of American Kestrel Nestling Development 22 day old nestlings (Cover image: 29 day old nestlings) 4

20 Contents Introduction 6 Methods 7 General Observations 8 At-a-Glance Kestrel Development Chart 9 Photographic Atlas Hatching-Day Day Day Day Day Day Day Day Day Post Day Conclusions and Acknowledgments 33 References 34 Key Terms 35 5

21 THE TIMELINE PROJECT Photographic Atlas of American Kestrel Nestling Development Introduction American Kestrels (Falco sparverius) are small, sexually dimorphic falcons sporting a distribution range in our region from Canada to Florida, with a few individuals being year-round inhabitants. After a largely monogamous courtship period ending in late March, female kestrels lay clutches of four to six eggs, with roughly one egg laid per day. Eggs hatch following an incubation period of about twenty-eight days. Once hatched, young nestlings develop to the point of fledging within a period of thirty to thirty-six days (personal observations). Literature references concerning kestrel development during this monthlong period however are scarce. The best guide in circulation that can aid in determining the age and gender of kestrel nestlings is that of Griggs and Steenhof (1993). The primary goal of this project is to clearly illustrate, through colored digital photographs and key measurements and observations, the developmental stages of typical male and female nestlings from time of hatching until fledging. Kestrel developmental variation previously has been correlated with numerous factors including: nestling gender, brood size, asynchronous hatching, parental conditioning (nutrition), and environmental effects (Gard and Bird 1992; Anderson 1993; Weibe and Bortolotti 1995; Dawson and Bortolotti 2000; Massemin et al. 2002). Due to these factors, our focus is to establish ranges of nestling development, which would take into consideration these sources of variation. As an auxiliary focus, this project hopes to aid field researchers working with the American Kestrels to provide an accurate at-a-glance method of aging young nestlings, and to better plan nest box visits for nestling banding. We hope that such a guide will be significant to those interested in Kestrel conservation, as well as those with a general interest in these falcons. 6

22 Methods Observations were taken on a study area near Hawk Mountain Sanctuary. The study area consists of 1500 square kilometers of open, sparsely wooded fields and farmland in Eastern Pennsylvania. Our site is 30 kilometers north of Reading and 30 kilometers west of Allentown, and includes portions of Berks, Lehigh, and Schuylkill counties. This location has been the site of several kestrel studies in the past (Rohrbaugh and Yahner 1997, Valdez et al. 2000). A conservation program, established by Hawk Mountain, has been maintained by Sanctuary staff and volunteers since 1987 (Rusbuldt et al. 2006). Kestrel broods used in this study nested in established nest boxes mounted to trees and, in a few cases, utility poles. Our study consisted of measuring and photographing nestlings from primarily three broods during June and early July of summer These three broods were visited on the day when the first nestling hatched until all of the nestlings fledged from the nest box. This provided us with an accurate starting age from which to base our subsequent measurements. These three broods comprised the entirety of our developmental measurement data, and supplied a total of 14 nestlings for our sample (with brood sizes of 4, 5, and 5 nestlings, respectively). Photographs were taken of other nestlings at different nest box locations, and their estimated ages were then determined using our observed data. Once the three study broods were identified and photographed at hatching, visits were made during early morning (9am-10:30am) every two days for additional photography. No measurements were taken prior to Day 9 due to a lack of significant primary feather sheaths. For each visit from Day 9 onward, in addition to the digital photographs, we measured with Vernier calipers the length of the fifth primary sheath, and after feather eruption the length of the fifth primary feather. Measurements were always taken from the left wing. Visual qualitative observations (color, feather patterns, gender determination) were noted on the nestlings appearance each visit. Nestlings were replaced in their nest boxes immediately following measurement sessions, which lasted approximately 15 minutes each. 7

23 General Observations Though our observations have conferred generalized indicators of nestling feather development, it must be stressed that variation is undoubtedly present. Variability factors such as brood size, hatching order, parental provisioning, nestling gender, and environment all readily affect nestling development, thus making a definitive description nigh impossible. The recommended developmental indicators outlined below pool our observations to account for some of this variability. Distinct observations of feather-sheath development are scarcely noticed until Day Prior to this time, nestlings are quite small and completely covered with white down. Nestling size increases during this period, but is difficult to quantify due to the variability factors previously mentioned. Monitoring these nest boxes led us to conclude that aging young during this period is difficult and requires judgment. Accurate aging during this period increases with the observer s experience, as a gestalt is developed. Once nestlings have reached Day 9-10, key characteristics begin to present themselves. Most notably, feather sheaths begin to appear on the wings and shoulders at this age. Size continues to increase, but the nestlings are still completely down-covered. Patches of colored feathers may appear on the breast at Day 11-12, while the primary sheaths lengthen. Tail feather development is still absent at this stage. As early as Day 13-14, the primary flight feathers may begin to erupt, though they will be quite small if present. Gender determinations may or may not be able to be made on the basis of primary feather color if they have emerged. At this age tail, flank, and back feathers also begin to erupt. By Day 15-16, gender can be determined with certainty based on typical adult feather coloring and patterning on wings and breast feathers. Back and flank feather density increase as the tail and primaries lengthen. From Day onward, size continually increases as the nestlings become more developed. Gender-specific tail banding as well as breast feather patterning emerges around Day The percentage of bodily down steadily decreases as body feathers lengthen and overall feather density increases. Developmental rates of males and females are roughly equal until Day During this last week of kestrel development prior to fledging, our observations show that male nestling feather development and size increase slightly more rapidly than females. 8

24 At-A-Glance Kestrel Development Chart Nestling Age Day 1-8 Day 9-10 Day Day Day Day Day Day Day > 25 Days Sheath-Primary #5 Length Not present Sheaths, if present: cm (Mean: 0.82cm) Sheaths: cm (Mean: 1.14cm) Sheaths: cm (Mean: 1.67cm) Primaries, if present: < 0.5cm (Mean: 0.28cm) Primaries: cm (Mean: 0.48cm) Primaries: cm (Mean: 1.53cm) Primaries: cm (Mean: 2.43cm) Primaries: cm (Mean: 3.71cm) Primaries: cm (Mean: 5.75cm) Not measured Characteristic Observations Small size, increases by day Sheaths absent, body covered in patchy white down Difficult to age accurately (Day 4-8) Size increases, down covered Primary sheaths and a few scapular sheaths may be present Sheaths developed, distinct Colored feathers (~10%) appear on breast sides No tail feathers Primaries begin to develop Tail, flank, & back feathers erupt Auriculars emerge and darken Possible to determine sex Feathers on back/shoulder & flank thicken Auriculars & primaries lengthen Sex is determinable Down prominent on head, lower back, & ~90% of wings Back & shoulders ~85% feathered Tail lengthens significantly (White terminal band of males/dark band of females is visible) Breast ~50-60% developed (Male spots/female streaking) Crown feathers emerge Entire back almost fully developed ~95% Upper wings ~20-30% down remaining Tail lengthens (Females gain 2 nd band) Breast ~80% full Crown thickens, little down remains on head Down present ~10-15% upper wings & crown (Females have more than males) Entire back 100% feathered Breast ~95% feathered on males (Some down remains on females ~10%) Tail feathers lengthen (5 bands for females) Males appear more fully feathered, some bodily down remains on females (lower breast, upper wing, lower back) Body size increases as fledging approaches Resemble miniature adult Kestrels 9

25 Photographic Atlas Recently hatched nestlings Hatch-Day 8: From the time of hatching until more than one week s time, nestlings are very small and are completely covered in white, patchy down. Size increases while down density decreases throughout this first week, although primary feather sheaths are completely absent at this stage. Lack of sheaths results in no clear indicator of development, making aging determinations in this range very difficult, especially from the age of Day 4 to 8. Aging in this range relies heavily on personal judgment and experience. When aging nestlings in this range, it is recommended to recheck the box within seven days to more accurately age nestlings. 10

26 One day old nestling with egg of sibling Two day old nestlings 11

27 Five day old nestling Seven day old nestlings 12

28 A Ten day old nestlings Day 9-Day 10: Early into the second week of nestling life, very little developmental indicators are present. Though nestling size increases, they remain fully down covered. At this stage primary and back feather sheaths may be exposed, though small. If sheaths are present A, they can range from 0.6 to 1.0cm (Mean: 0.82cm). 13

29 B Eleven day old nestlings Day 11-Day 12: B During this stage of kestrel development, the prominent indicator of age is the exposure of the primary feather sheaths B,C, which are very distinct. Sheaths range at this age from 0.7 to 1.6cm (Mean: 1.14cm). Patches of colored feathers begin to emerge on the exterior regions of the breast. Nestlings backs remain completely down covered and tail feathers are absent. 14

30 C Eleven day old Male nestling Twelve day old Female nestling 15

31 F Fourteen day old Female nestling Day 13-Day 14: After two weeks of growth, sheaths are still the dominant anatomical feature with ranges from 1.4 to 2.0cm (Mean: 1.67cm). However, at this stage of development, other indicators of age also begin to emerge. Most notably, primary feathers begin to erupt after approximately two weeks, although at lengths less than 0.5cm (Mean: 0.28cm) D. Feathers on the tail E, flank F, and back also begin to erupt in this period. The feathers covering the ear opening (auriculars) begin to darken at this age, and constitute another distinctive indicator G. Depending on the brood, nestling gender may also be determined at this point by observation of blue-gray coloration on the male wing, and brown and black pigmentation on the female. 16

32 D G Fourteen day old nestlings E 17

33 I Sixteen day old Female nestling Day 15-Day 16: By the age of fifteen or sixteen days, nestling gender can be determined accurately as primary feathers have emerged (Range: 0.2 to 0.9cm, Mean: 0.48cm) and bear the gender-specific pigmentation seen in adult birds (blue-gray coloration of wings on males, brown and black on females) H. Exterior flank feathers increase in density, highlighting the spotting patterns of the males and the barring pattern of the females I. Back feathers have erupted rather significantly (~30% feathered) J, and increase in density across much of the dorsal region. 18

34 J H Sixteen day old Male nestling 19

35 O N Eighteen day old Male nestling Day 17-Day 18: At this stage, primary feathers continue to lengthen, ranging from 1.1 to 2.1cm (Mean: 1.53cm) K. Back feathers have filled in almost completely, with female alternating brown-and-black barring and the male rufous-and-black barring L. Flank and breast feathers continue to increase in density as they develop inwards from the exterior M. Male breast spotting and female barring increase in intensity. Tail feathers have increased significantly, and terminal banding appears (a white band in males, a dark band in females) N. Head remains largely down covered, though crown feathers do begin to emerge O. 20

36 M Eighteen day old Female nestlings K 21

37 L Eighteen day old Male nestling 22

38 R Twenty day old Female nestling Day 19-Day 20: In this time period, primary feathers continue to grow (Range: 2.0 to 3.2cm, Mean: 2.43cm) P, as do the back Q and breast feathers R, with these areas almost completely feather covered. Tail feathers lengthen, with females showing a second brown band S. Crown feather density increases, with little down remaining on the head. 23

39 Q Twenty day old nestlings S P 24

40 Twenty-two day old Female nestling Day 21-Day 22: At three weeks of age, primary feathers range in length from 3.3 to 4.5cm (Mean: 3.71cm) T. Feathers on the back are almost fully developed (~95-100% feathered) U, as are breast feathers on males. Female breast feathering, it appears, is at a lower percentage than males. On a whole, more bodily down remains on females than males. Rufous patch on the males crown is distinctly visible by this age V. The tail has lengthened, and male banding pattern (white terminal, black midline, rufous base) W is present but no new bands have developed on the female. Head banding pattern of alternating dark and light feathers (mustache, auricular, nape) is highly pronounced X. 25

41 T U Twenty-two day old nestlings V W 26

42 X Twenty-one day old Male nestling 27

43 Twenty-three day old nestlings Day 23-Day 25: Toward the end of the kestrels time in their nest boxes, primary feathers reach lengths of 5.2 to 6.2cm (Mean: 5.75cm) Y. At this age, kestrels appear as miniature adults. Males appear more fully feathered and have less observable down than females, who still have noticeable patches of down on their lower breast and back, as well as upper wings. Tail feathers lengthen significantly, with females having five alternating bands at this age Z. 28

44 Y Twenty-five day old nestlings Z 29

45 Twenty-five day old nestlings 30

46 Twenty-eight day old Male nestling Post Day 25: From Day 25, no new measurements or photographs were purposely taken. However, a few candid photographs were taken when we surprised the nestlings still in the boxes. Boxes were monitored over the next week for signs of fledging. During this pre-fledging period, kestrel size increased as nestlings came to fully resemble miniature adults. The fourteen nestlings we observed fledged as early as Day 30 and as late as after Day

47 32 Thirty-one day old Female nestlings

48 Conclusion Following and photographing the development of these young American Kestrels has been an interesting and rewarding experience. It was a wonderful feeling seeing these birds struggle to exist, to observe their survival through to fledging, and to have the chance to be part of the American Kestrel population in Berks County. We hope we have the opportunity to observe these fledglings on our study site again within the next few breeding seasons. Finally, we hope that you, the reader, enjoyed this reference guide, and join in the support and conservation of wildlife in Pennsylvania and beyond, and that you are encouraged to learn more about the American Kestrel. Acknowledgments We offer our thanks to Jim s wife Kelly, for her help and support throughout the project, and especially for her artistic photography, and to Bob and Sue Robertson, our friends and fellow field biologists. We would especially like to thank Keith Bildstein at Hawk Mountain Sanctuary for his editing and technical support. Additional thanks go to Bill Thorne for his help in reviewing our data and Rev. Dr. Richard Rusbuldt for editing. Finally, we send our gratitude to Mark Rittwage at Complete Graphics Inc. for his design advice and layout editing, and to Alvernia College s Dr. Carrie Fitzpatrick for text editing and guidance throughout the publication process and Michele Spotts for photograph editing and cover design. Twelve day old Female nestling 33

49 References Anderson, David Prey size influences female competitive dominance in nestling American Kestrels. Ecology 74: Dawson, R.D. and G.R. Bortolotti Reproductive success of American kestrels: The role of prey abundance and weather. The Condor 102: Gard, N.W. and D.M. Bird Nestling growth and fledging success in manipulated American Kestrel broods. Canadian Journal of Zoology 70: Griggs, G.R. and K. Steenhof Photographic guide for aging nestling American Kestrels. U.S. Department of the Interior, Bureau of Land Management, Boise, Idaho. Massemin, S., E. Korpimäki, V. Pöyri, and T. Zorn Influence of hatching order on growth rate and resting metabolism of kestrel nestlings. Journal of Avian Biology 33: Rohrbaugh, R. and R. Yahner Effects of macrohabitat and microhabitat on nest-box use and nesting success of American Kestrels. The Wilson Bulletin 109: Rusbuldt, J.J., J.R. Klucsarits, S. Robertson, and B. Robertson Reproductive success of American Kestrels using nest boxes in Eastern Pennsylvania, Pennsylvania Birds 20: Valdez, U., S. Robertson, B. Robertson, and K.L. Bildstein Nest box use by American Kestrels (Falco sparverius) and European Starlings (Sturnus vulgaris) in Eastern Pennsylvania. Pennsylvania Birds 14: Weibe, K. L and G. R. Bortolloti Food-dependant benefits of hatching asynchrony in American kestrels Falco sparverius. Behavioral Ecology and Sociobiology. 36:

50 Key Terms Asynchronous Hatching Occurs in birds that begin incubation of eggs prior to laying the last egg of the clutch. Auricular Feathers Feathers that cover the opening of a bird s ear. Barring Pattern Alternating light and dark feather bands in the plumage of American Kestrels. Crown Feathers Feathers that cover the top of the head, specifically behind the eyes. Primary Feathers Long or large stiff feathers on the last section of a bird s wing. Primaries are numbered from the innermost to outermost. Primary Sheaths Keratinized material that surrounds developing feathers. Sheaths are numbered in the same manner as feathers. Scapular Feathers Grouping of feathers on the shoulders and sides of the back. Sexual Dimorphism Visual differences in the plumage of the sexes of a species. Spotting Pattern Dark circular feather patches adorning the breast and flank of male American Kestrels. Terminal Band The culminating band of American Kestrel tail feathers. 35

51 About the Authors James Klucsarits is a volunteer researcher with Hawk Mountain Sanctuary and has been actively studying and banding American Kestrels since Together with fellow volunteers and interested students, he has been monitoring the long-term reproductive success of kestrels in Eastern Pennsylvania. He also is a professor of biology at Alvernia College and is extremely interested in the conservation of all avian species. At the time of this writing, Joshua Rusbuldt is a junior at Alvernia College majoring in biochemistry. He began working with Klucsarits during his freshman year and has been an active volunteer in the kestrel conservation effort ever since. With an avid interest in all things science, working with these falcons has helped shift his focus from a textbook idealism to the natural workings of a much larger magnitude. Authors James R. Klucsarits (left) and Joshua J. Rusbuldt (right) 36