western painted turtle monitoring at alaksen national wildlife … · 2011. 4. 28. · western...

Western Painted Turtle Monitoring at Alaksen National Wildlife Area (NWA)

Final Report – March 2011

Prepared by

Vanessa Kilburn and Aimee Mitchell

of the

The South Coast Western Painted Turtle Recovery Project

For

Stephen Hureau, Canadian Wildlife Service, Delta, BC

Western Painted Turtle Monitoring at Alaksen NWA: Summary Report December 2010


Table of Contents Introduction ................................................................................................................................................................4

Project Rationale ....................................................................................................................................................4

Species Description ................................................................................................................................................4

Site Description ......................................................................................................................................................5

Overview .............................................................................................................................................................5

Aquatic Habitat ...................................................................................................................................................6

Nesting Habitat ...................................................................................................................................................6

Landscape Composition ......................................................................................................................................7

Site-Specific Threats ...............................................................................................................................................7

Agriculture and pollution ....................................................................................................................................7

Invasive species ..................................................................................................................................................7

Road-related threats and collection ...................................................................................................................7

Methods .....................................................................................................................................................................8

Surveying, Monitoring, Tracking, and Habitat Characterization ............................................................................8

Data Analysis of Habitat Characteristics .................................................................................................................8

Results ........................................................................................................................................................................8

Surveying ................................................................................................................................................................8

Monitoring ..............................................................................................................................................................9

Radio-Tracking and Movements .......................................................................................................................... 11

Habitat Plot Characterization .............................................................................................................................. 11

Characterization of Overwintering Habitat ......................................................................................................... 14

Discussion ................................................................................................................................................................ 14

Population Status ................................................................................................................................................ 14

Habitat Use and Movement ................................................................................................................................ 15

Management Recommendations ............................................................................................................................ 15

Population Augmentation ................................................................................................................................... 15

Invasive Species Removal .................................................................................................................................... 16

Habitat Enhancement .......................................................................................................................................... 16

Additional Research ............................................................................................................................................. 16

Outreach and education ...................................................................................................................................... 16

References ............................................................................................................................................................... 17



List of Figures Figure 1: Red-eared Slider (left) and Western Painted Turtle (right) basking on a log at Alaksen NWA. ..................4

Figure 2: Alaksen National Wildlife Area (NWA) overview map. ...............................................................................5

Figure 3: Aquatic habitat at Alaksen NWA. ................................................................................................................6

Figure 4: Potential Nesting Area (PNA) locations (outlined in yellow) for turtles at Alaksen NWA. .........................6

Figure 5: Radio-marked Western Painted Turtle at Alaksen NWA. ............................................................................9

Figure 6: Locations of traps (white symbols) and capture/re-capture locations for Western Painted Turtle at

Alaksen NWA summer 2010.. .................................................................................................................................. 10

Figure 8: Western Painted Turtle habitat plots at Alaksen NWA. ........................................................................... 12

Figure 9: Western Painted Turtle overwintering location at Alaksen NWA. ........................................................... 14

List of Tables Table 1. Western Painted Turtle Occurrence Surveys at Alaksen NWA ..........................……………………………………..9

Table 2. Summary of Trapping Effort and Results at Alaksen NWA...........…………………………………………………………10

Table 3. Habitat characteristics and analyses at Western Painted Turtle tracking and random locations............13

List of Appendices Appendix A. Western Painted Turtle (Chrysemys picta bellii) Surveying and Monitoring Protocols……………….….19

Appendix B. Western Painted Turtle Blood Sampling Protocol…………………………………………………………………………..32



Figure 1: Red-eared Slider (left) and Western Painted Turtle

(right) basking on a log at Alaksen NWA.

Introduction

Project Rationale

The Pacific Coast Population of the Western Painted Turtle is considered to be Endangered by COSEWIC, on

SARA Schedule 1, and is provincially red-listed. To date there are 61 known occupied sites for this species on the

south coast [1], disjunctly distributed throughout the Lower Mainland, on the Sunshine Coast north to Powell

River, throughout southern and central Vancouver Island, and on some Gulf Islands, including Salt Spring Island

and Texada Island. The Western Painted Turtle is more abundantly distributed in the low-lying Rocky Mountain

Trench and other lowlands of the southern interior of BC; this population, known as the Intermountain-Rocky

Mountain Population, is considered distinct from the Pacific Coast Population and is listed as Special Concern by

COSEWIC and is provincially blue-listed [1]. Across its range, the Western Painted Turtle’s preferred aquatic

habitat is slow-moving, shallow water bodies including small lakes, lagoons, marshes, ponds, and sloughs with

ample woody debris and aquatic vegetation [2]. Upland terrestrial habitat is required for nesting, and like other

species of freshwater turtles, the Western Painted Turtle prefers open, south-facing areas with sandy loose

substrate [2].

In 2009, Western Painted Turtles were identified for the first time at Alaksen National Wildlife Area (NWA), and

this site became one of the few known Western Painted Turtle occupied sites in the Lower Mainland. Because

this site is located on federal land and this species is federally listed, it was deemed of importance to initiate a

research and monitoring program for this population of Western Painted Turtles. In July 2010, a mark-recapture

and radio-telemetry program was initiated to enable researchers to investigate seasonal movement and habitat

use of this species at Alaksen NWA. This document contains the results of this study, including a summary of the

habitat types most commonly used by turtles, maps showing seasonal movement patterns, and management

recommendations for the turtle population at Alaksen NWA.

Species Description

The Western Painted Turtle (Chrysemys picta bellii)

is the only native freshwater turtle species in B.C.

However, in many sites across its range it co-

occurs with the introduced Red-eared Slider

(Trachemys scripta elegans) and may be confused

with this species. The Red-eared Slider is

distinguishable from the Western Painted Turtle

by the presence of a red ear mark on the sides of

the head (sometimes absent in old individuals), a

higher domed carapace that is slightly keeled

along the length, and a yellow plastron and

marginals with dark abstract patterning. Sliders

are usually larger, but differences are not

necessarily obvious unless the two species are

observed alongside one another (as in Figure 1).



Figure 2: Alaksen National Wildlife Area (NWA) overview map. Top right corner inset shows monitoring area, labelled as 1

through 5 (from bottom to top); areas 1 through 4 are lagoons while area 5 is a small marsh.

Both species have irregular yellow lines on the neck but they are thinner and denser on the Western Painted

Turtle. Males of both species have longer claws on the forefeet and wider tails compared to females [3].

Site Description

Overview

The Alaksen NWA is located on Westham Island in the Municipality of Delta, 40 km south of Vancouver, in the

Fraser River estuary (Fig. 2). In total, the site comprises 586 ha of protected area managed for migratory birds;

300 ha of this site is Alaksen NWA, and the remaining area is the adjacent George C. Reifel Migratory Bird

Sanctuary [4]. Land tenure for the area is divided between the federal government (approx. 53%) and the

provincial government (approx. 47%) [4]. This area has been officially protected under the Canada Wildlife Act

since 1973, and in 1987 the site was designated a Wetland of International Significance under the Ramsar

Convention, the 9th

designated Canadian Ramsar site [4]. However, the surrounding fields are leased to farmers

and used for agriculture. Our study area is shown in the inset of Figure 2. For the purpose of monitoring, water

bodies used by turtles were labelled Lagoon or Wetland 1 through 5.



Figure 3: Aquatic habitat at Alaksen NWA.

Aquatic Habitat

Under the Ramsar Classification System, the

wetlands at Alaksen NWA are classified as

permanent freshwater ponds, marshes and swamps

[4]. These water bodies are generally at most 2-3 m

deep. The bottom substrate in most areas is

sand/gravel covered by a layer of organic detritus,

although in other areas the bottom is very

muddy/clayey. There is a great deal of submergent

and emergent woody debris which serves as critical

basking habitat for turtles, and important perching

habitat for birds and cover habitat for fish (Figure 3).

However, emergent, submergent and floating-

leaved vegetation, which adult turtles prefer to

forage on, is limited. These wetlands seldom freeze

over for more than three weeks in the winter [4].

Nesting Habitat

There is some surrounding upland habitat within 50 m from the water‘s edge of Lagoon 1 that may serve as

nesting habitat for turtles, and there is evidence that Western Painted Turtles as well as Red-eared Sliders have

been attempting to nest in this area (Fig. 4). The success of these attempts remains unknown. In spring 2009,

several Canadian Wildlife Service employees reported seeing both species walking on land in the area beside the

visitor parking lot (PNA1), and the caretaker of the site has seen turtles laying nests on his lawn (PNA3) in the

past but was not able to confirm whether these attempts were by Sliders or Painted turtles. In spring 2009

surveys, several test holes were found in PNA1, but no occupied nests were identified (Fig. 4). This area is

composed of sandy substrate and is an open south-facing area, however it is becoming overgrown with invasive

vegetation (mainly blackberry).

Figure 4: Potential Nesting Area (PNA) locations (outlined in yellow) for turtles at Alaksen NWA. Yellow markers

are locations where Western Painted Turtles were observed and Red markers indicate Red-eared Slider

observations.



Landscape Composition

Landscape types present at this site include cultivated agricultural fields, freshwater and brackish tidal marsh

wetlands, woodland or grassland, and built-up or barren shore flats. Landscape composition is similar in all three

upland buffer zones (50 m, 200 m and 350 m surrounding the water’s edge) with forest, agricultural fields, and

fresh or saltwater systems making up the majority of composition in each zone. Road (paved and gravel) and

path density is low in each zone, and development is also minimal and localized to two small areas supporting

Canadian Wildlife Service (CWS) office buildings and Reifel Bird Sanctuary visitor buildings.

Site-Specific Threats

Threats to Western Painted Turtles at Alaksen NWA are minimal owing to this site‘s protected status, however

there are a few potential threats that may have an impact on the overall health of turtle populations. Identified

threats to this population include water pollution as a result of agricultural practices, invasive species, and road-

related mortality and collection. These threats are described in more detail in the following sections.

Agriculture and pollution

Pollution by toxic substances from adjacent agricultural areas is a potential threat to turtles at this site, as

pollution from herbicides and pesticides is known to be high in the Lower Fraser River Valley [5,6]. CWS has

recently conducted water quality sampling and analyses at Alaksen NWA, and has found that residual pesticide

levels are fairly high in some areas, at least for certain pesticides [7].Chemical contamination in water resulting

from agricultural effluents can affect the reproductive output of freshwater turtles, result in chromosomal

damage causing developmental abnormalities, malformations and cancerous tumours, and cause endocrine

disruption resulting in skewed sex ratios [8,9,10,11]. Additionally, agricultural fields could represent high-risk

areas for overlanding turtles, especially if the fields are regularly tilled or otherwise mechanically disturbed. The

results from this study, along with inspections for abnormalities in individual turtles, will be used to determine

the potential impacts to turtles from agricultural pollution.

Invasive species

Invasive species are likely to represent a threat to turtles at this site. Invasive plants in upland terrestrial areas

tend to choke out suitable nesting habitat [12], and invasive animal species in aquatic systems may act as

resource competitors and disease agents (Red-eared Sliders) [13,14] or prey on young turtles (Carp and Bullfrogs

[15]). Red-eared Sliders, Carp, Bullfrogs and Green Frogs have all been confirmed at this site. Impacts from

invasive turtles and frogs at this site are unlikely to be a large threat to adult turtles because there is extensive

essential aquatic habitat. However, these species could have a significant impact on hatchling or juvenile turtles.

Invasive plant species, such as Himalayan Blackberry (Rubus discolour), in upland nesting areas may however

present more of a threat currently.

Road-related threats and collection

Finally, roads and trails are known to pose severe threats to turtle populations, acting most severely as direct

mortality factors (roads)[16] but also by facilitating access to turtles resulting in opportunistic collection

(trails)[17]. However, road density at Alaksen NWA is low and roads are limitedly used by CWS employees and

park visitors. Both the Alaksen NWA and Reifel Bird Sanctuary are open to the public and contain a network of

walking trails, but collection of turtles in this area is unlikely due to constant on-site surveillance. The biggest

concern at this site with respect to road-related mortality is the visitor parking lot located immediately adjacent



to the most well-used nesting site, PNA1 (Fig. 2). Visitors unaware of the possibility of turtles on land in this area

could easily back their vehicles over a turtle, and certainly there have been many reports of turtles walking

through this parking lot during the spring and summer months, when the parking lot is heavily used.

Methods

Surveying, Monitoring, Tracking, and Habitat Characterization

A combination of methods was used in this study for inventorying and monitoring turtles at Alaksen NWA.

Detailed methodology appears in Appendix A in the form of protocols. Briefly, population inventories were

conducted using basking surveys by land and kayak during June 2010 when turtles are basking and most visible.

Ten baited hoop traps were deployed for a 3-week period from mid-July through mid-August to catch turtles. All

captured turtles were measured, weighed, sexed, and marked with individual shell notch codes for future

identification. The Western Painted Turtles was fitted with a radio-transmitter (Holohil AI-2F, 30 g, Frequency

148.519) and radio-tracked intermittently over a 4-month period, from late August to early November 2010, and

then one final time on March 7th

, 2011. Additionally, we collected a caudal blood sample from this individual for

future population genetic analysis, following a protocol developed for the Ministry of Environment by Elinor

Hughes (2010; see Appendix B).

During each tracking event, habitat was assessed at the turtle’s identified location; habitat measurements taken

included water depth, water temperature at various depths, water quality (pH and conductivity), air

temperature and basking log temperature, and habitat features were quantified (aquatic vegetation, woody

debris, composition of bottom substrate). We also assessed habitat at randomly generated points to compare

habitat characteristics at the turtle’s chosen location to a random location in the wetland complex. The

surveying and monitoring protocols have been prepared so that Canadian Wildlife Service biologists may

continue surveying and monitoring efforts at this site in the future. The choice of habitat characteristics to

measure was based on Marchand and Litvaitis (2004).

Data Analysis of Habitat Characteristics

To compare habitat characteristics between the turtle’s chosen location and random locations within the study

lagoons at Alaksen NWA, we used a paired t-test to test for differences in continuous variables (e.g., mean

temperature, depth, conductivity, pH) and Chi-squared Contingency Analysis to test for differences between all

categorical variables (e.g., emergent/submergent vegetation and woody debris, habitat type, and substrate

type). Temperatures were separated into summer (July-August) and fall-winter (September-November and

March) periods. For many of the categorical variables, fields had less than 5 variables, and thus had to be

combined in order to test for differences using Chi-square tests. For emergent/submergent vegetation and

woody debris, the “sparse” and “scattered” categories were combined, and the “common” and “abundant”

categories were combined. Three categories for substrate type were created; 1) sand, 2) sand/detritus with

sand/muck, and 3) muck/detritus, muck/clay and/or mud.

Results

Surveying

Three turtle occurrence surveys were conducted at the Alaksen NWA in 2010 for a total of 6 search effort hours;

Western Painted Turtles, Red-eared Sliders, or both were encountered on all three surveys (Table 1). The



maximum number of individual Western Painted Turtles observed during a single survey session was two, while

as many as six Red-eared Slider individuals were identified during a single survey. Occurrence surveys were also

conducted at the adjacent G.C. Reifel Bird Sanctuary on two occasions (4 Search Effort Hours) but turtles were

not observed during either of these surveys. There are however anecdotal reports of Red-eared Sliders at this

site, however Western Painted Turtles have not been reported from the Reifel Bird Sanctuary.

Table 2: Western Painted Turtle Occurrence Surveys at Alaksen NWA in Summer 2010.

Site Survey # DateSearch Effort

HoursWPT (# Ind.)

UTM (10 U WGS84)

ActivityOther Turtles

Species UTM (10 U

WGS84)Activity

2487602 5438660

Basking on log

1487546 5438661

Basking on log w/ slider - same PT as earlier?

1487546 5438661

Basking on log with slider 1 Red-eared Slider487546 5438661

Basking on log w/ PT

1487561 5438670

Basking on log 1 Red-eared Slider487463 5438849

Basking on log - northwest lagoon

1 Red-eared Slider487552 5438650

Basking on log


Basking on log


Basking on log

1 Unknown Turtle487460 5438601

Basking on log with above sliders - WPT?

1 Jun-04 2 0 N/A N/A 0 N/A N/A N/A

2 Jun-17 2 0 N/A N/A 0 N/A N/A N/A

2

3 Aug-11 1.5

Alaksen Wildlife Refuge

1 Jun-04 2.5 1 Red-eared Slider487546 5438661

Basking on log w/ PT

Reifel Bird Sanctuary

? N/A N/A

2 Jun-17

Nesting surveys were conducted at this site in 2009 and then again in 2010. No turtles were seen on land during

either years, but test holes were located in one area in 2009 where there have been reports of turtles on land.

No occupied nests could be confirmed in this location at this time; however a temperature/humidity data logger

was installed here because of the high possibility of occupied nests in this area. In 2010, no test holes were

located in any of the identified potential nesting areas and there were no reports of turtles on land.

Monitoring

From the 14th

of July to the 6th

of August, 2010, we

trapped turtles with an average of 219.25 trap hours

per trap (Range 110.17- 257.58) (Table 2). Trapping

efforts resulted in the capture of one Western

Painted Turtle (WPT) female that was subsequently

measured, marked and fit with a radio-trasmitter

(Table 2 and Figure 5). However, the appearance of

the turtle was suggestive of a Western and Southern

Painted Turtle hybrid ( i.e., red lines on neck and

limbs and on tail; radiating plastron marking not quite

to edges of plastron; plastron yellowish-orange;

light middorsal stripe). This individual was captured

on the 15th

of July, 2010 and was re-captured four

Figure 5: Radio-marked Western Painted Turtle at Alaksen

NWA.



A

Figure 6: Locations of traps (white symbols) and capture/re-capture locations for Western Painted Turtle at Alaksen NWA

summer 2010. A - Lagoon 1 trap locations and B - Lagoon 2 and 3 trap locations. The green symbol represents the initial trap

the Western Painted Turtle was captured in and red symbols represent the trap recapture locations for this individual.

B

times in 11 days in four separate traps (Fig. 6). Although one additional Western Painted Turtle was observed on

three other occasions near the traps in Lagoon 1, it was not captured. Nine Red-eared Sliders (RES) were also

captured, measured and marked, and two individuals were recaptured once. In addition to turtles, 28 Carp and

13 Catfish were caught in the hoop traps and released (Table 2).

Table 2. Summary of Trapping Effort and Results at Alaksen NWA Summer 2010.

Trap # Location# WPT captured /

recaptured

WPT markings captured / recaptured

# RES captured / recaptured

RES markings captured / recaptured

Other Species

captured (#)

Trapping Effort Hours

1 Lagoon 1 0 / 1 N/A / 5-1 0 / 0 N/A Carp (1) 258.37

2 Lagoon 1 1 / 15-1 (radio-

marked) / 5-16 / 0

5-1, 5-3 to 5-7 / N/A

Carp (1) 257.55

3 Lagoon 1 0 / 0 N/A 2 / 05-2, 5-23 /

N/ACarp (1) 257.58

4 Lagoon 1 0 / 1 N/A / 5-1 0 / 1 N/A / 5-2Carp (2),

Catfish (1)257.33

5 Lagoon 1 0 / 1 N/A / 5-1 0 / 0 N/A Carp (3) 257.25

6 Lagoon 1 0 / 0 N/A 0 / 0 N/A Carp (1) 257.17

7 Lagoon 2 0 / 0 N/A 1 / 1 4-2 / 5-1 Carp (3) 239.17

8 Lagoon 1 0 / 0 N/A 0 / 0 N/A Carp (10) 207.25

9 Lagoon 3 0 / 0 N/A 0 / 0 N/A Carp (4) 100.67

10 Lagoon 3 0 / 0 N/A 0 / 0 N/ACarp (2),

Catfish (12)100.17



Radio-Tracking and Movements

Thirty-nine tracking locations were acquired for the radio-marked turtle, including its original capture location

and subsequent recaptures from the 15th

of July 2010 to the 7th

of March 2011 (Fig. 7). Four intensive radio-

tracking sessions monitored daily summer movements. These movements averaged 97.8 m every 2 hours from

0800 to 2000 hours, with 582 m being the furthest distance recorded travelled in a 2-hour period. From the

turtle’s original trap location in July to its last tracking in early March, this turtle navigated a total of 2.0 km

(straight distance) of four sloughs and one wetland in the Wildlife Area beginning in a southern slough and

overwintering in a northern wetland (Fig. 7). The overwintering location was reached in late September.

Figure 7. Locations (green circles) of the radio-tracked female Western Painted Turtle and corresponding time-line at

Alaksen NWA between July 2010 and March 2011.

Habitat Plot Characterization

Habitat characteristics were recorded at 59 plots; 30 at tracked turtle locations and 29 at random locations (Fig.

8 and Table 3.). Analyses indicates this Western Painted Turtle overwhelmingly selected edge habitat (including

basking habitat), with a significantly higher portion (97%) of habitat plots conducted at tracked turtle locations

in this type of habitat. Random plots, on the other hand, were only in edge habitat 39% of the time. In

association with edge habitats, this turtle was also using shallow areas with an average depth of 92 cm,

Late-September- Mid-

March



significantly shallower than random locations (mean = 134 m). Locations selected by the tagged turtle were also

characterized as being shady areas with a higher percentage of cover, were more likely to have common and

abundant woody emergents and submergents and have a sandy substrate with some organics (detritus or muck)

rather than purely sand. No difference, in either summer or fall, was detected in any of the temperature, Ph or

conductivity measurements between tracking and random locations.

Figure 7: Western Painted Turtle habitat plots at Alaksen National Wildlife Area sampled late summer through fall

2010; Green circles represent plots at radio-marked turtle locations and red circles represent random plots.



Habitat Characteristics Tracking locations (n = 30)

Proportion Random locations (n = 29)

Proportion Statistic* Comments

Temperatures (oC) - summer / fall meansa

Air 22.5 / 17.5 N/A 21.9 / 17.0 N/A p = 0.73 / p = 0.62

No difference in summer or fall

Surface Water 23.5 / 16.6 N/A 23.7 / 16.3 N/A p = 0.39 / p = 0.59


Column Water 23.0 / 16.0 N/A 23.2 / 15.8 N/A p = 0.35 / p = 0.54


Bottom Water 22.9 / 16.0 N/A 22.9 / 15.8 N/A p = 0.47 / p = 0.54


Nearest bask 23.8 / 16.4 N/A 22.8 / 16.8 N/A p = 0.90 / p = 0.42


Conduct - meana 4.70 / 5.65 N/A 4.89 / 6.72 N/A p = 0.14 / p = 0.34


Ph - meana 9.12 / 8.01 N/A 9.14 / 8.15 N/A p = 0.33 / p = 0.31


Depth (cm) - mean 92.0 N/A 133.5 N/A p < 0.0001 Tracking locations are significantly

at shallower (<1m) depths

Aquatic habitat type # of points # of points Edge (ED) - (Basking

(BA) 29 (9 BA) 0.97 12 (1 BA) 0.41

p < 0.0001 Tracking locations are significantly

in Edge habitat Open water (OP) 1 0.03 17 0.59

Degree of cover # of points # of points Clear (CE) 14 0.47 21 0.70

p = 0.13 Tracking locations are more often

located in Shady areas but not significantly Shady (SH) 16 0.53 9 0.30

% Shady - mean 65.0 N/A 48.1 N/A p = 0.96 Tracking locations in shady areas have a higher degree of cover but

not significantly Emergents/ Submergentsb - 0 / 1 / 2 / 3 # of points # of points

Woody Emergents 4 / 3 / 15 / 8 .13 / .10 / .50 / .27 15 / 3 / 8 / 3 .52 / .10 / .28 / .10 p < 0.01 Tracking locations are significantly more often in areas with common and abundant woody emergents

Woody Submergents 9 / 1 / 15 / 5 .30 / .03 / .50 / .17 17 / 1 / 9 / 2 .59 / .03 / .31 / .07 p < 0.01 Tracking locations are significantly more often in areas with common

and abundant woody subemergents

Emergent Vegetation 21 / 3 / 4 / 2 .70/ .10 / .13 / .07 28 / 1 / 0 / 0 .96 / .04 / .00 / .00 p = 0.01 Random locations are more likely

to be in areas with sparse and scattered emergent vegetation

Submergent Vegetation

27 / 3 / 0 / 0 .90 / .10 / .00 / .00 28 / 1 / 0 / 0 .97 / .03 / .00 / .00 N/A Submergent vegetation at all

locations was sparse and scattered - no difference

Water visibility c - 0 / 1 / 2 30 / 0 / 0 100 / 0 / 0 29 / 0 / 0 100 / 0 / 0 N/A Water visibility in all waterbodies

was opaque - no difference

Substrated # of points # of points Sand 8 0.27 18 0.62

p < 0.0001

A greater proporation of random points were in purely sandy areas than any other category. Tracking points more frequently occurred in

sandy areas with some organics (detritus or muck) or in areas with

purely organics

Sand/detritus 7 0.23 0 0.00

Sand/muck 4 0.13 0 0.00

Muck/detritus 7 0.23 2 0.07

Muck/clay 3 0.10 7 0.24

Clay and/or mud 1 0.04 2 0.07

aTemperatures separated into summer (July-August) and fall (September-November) periods

bEmergents / Submergents: 0 - sparse, 1- scattered, 2 - common, 3 – abundant within 2 m. Categories were combined into two; 0 - sparse with 1- scattered; AND 2-common with 3-abundant for analyses b/c Chi-square suspect with more than 20% of fields with less than 5

cWater visibility : 0 - completely opaque, 1 - 50% transparent, and 2- clear

dSubstrate - Categories were combined into three; 1) sand, 2) sand/detritus with sand/muck, AND 3) muck/detritus with muck/clay and clay and/or mud for analyses b/c Chi square suspect with more than 20% of fields with less than 5

Table 3: Habitat characteristics and analyses at Western Painted Turtle tracking and random locations at Alaksen

National Wildlife Area, sampled late summer 2010 through to winter 2011.



Characterization of Overwintering Habitat

The habitat at the chosen overwintering location was very different from all other aquatic habitat at Alaksen

NWA. Firstly, there was ample aquatic vegetation, both emergent (mostly cattails) and submergent, compared

to at all other areas of the site. The wetland is small and shallow with stagnant water and has abundant

emergent and submerged woody debris (Fig. 9). The bottom substrate is organic in nature with a deep substrate

profile, in contrast to the bottom of the vast majority of the sloughs at this site, which are composed of sandy

substrate with a layer of mud and clay. The riparian area consists of deciduous trees, blackberry and spirea, with

overhanging branches. During all winter tracking events (two in November and one in March), the Western

Painted Turtle was located in shallow (ca. 50 cm) edge habitat with overhanging vegetation. A distance of 20 m

separated the November locations from the March location. Although we did not detect movement by the turtle

in November, in March the turtle appeared to be moving slightly along the bottom as evidenced by changes in

location of the strongest tracking signal. The water temperature on this day was 6 °C.

Discussion

Population Status

Only two Western Painted Turtles were observed at one time at this site. No juveniles were captured during the

trapping session, despite the fact that they tend to be easier to catch in the fish-baited traps than adults due to

their more carnivorous foraging habits (personal observations from the authors). Additionally, the tagged

Painted Turtle shows characteristics of both a Western and Southern Painted Turtle, and thus the origin of this

individual and even population cannot be determined. However, individual variations in colour patterns have

been noted in other areas for this species, so it is also possible that this turtle is a true Western Painted Turtle

with colour and pattern variation, and not a hybrid.

Figure 8: Western Painted Turtle overwintering location at Alaksen National Wildlife Area. The left photo is the wetland in

September and the right photo is the wetland in November 2010. Turtle overwintering location is in bottom left corner of

wetland where some ice on the water surface has melted.



Red-eared Sliders have obviously been released into this site, so there is a possibility that the individual tagged,

and even the additional individual that has been observed, are released pets. There have been debates over

whether the Pacific Coast Western Painted Turtle population is native or introduced, however COSEWIC has

declared this Population native [15]. There is still a possibility however that some local populations are

introduced. A blood sample was taken from the tagged individual for future population genetic analysis, which

will aid in our understanding of the origin of Western Painted Turtle populations on the south coast, including

the population at Alaksen NWA.

Habitat Use and Movement

Results indicate that Western Painted Turtles at this site are utilizing the abundance of habitat available for

foraging, basking, and overwintering. Across the North American range of the Painted Turtle, preferred habitat

consists of shallow, slow moving, stagnant water with ample emergent and floating vegetation [16, 17, 18]. In

southeastern North America, abundance of Eastern Painted Turtles is positively correlated with abundant

aquatic vegetation and bottom substrates composed of organic matter like decaying plants; organic bottom

substrates provide turtles with suitable overwintering habitat, and aquatic vegetation provides foraging

opportunities for turtles [21]. However, foraging sites are lacking at Alaksen NWA; although adult freshwater

turtles are omnivorous, they do tend toward herbivory [22], and except for the small wetland on the northern

edge of the property where the tagged individual overwintered, aquatic vegetation is almost completely lacking

at this site.

The overwintering habitat chosen by this tagged turtle is similar to overwintering habitat choice reported for

turtles in sites outside of B.C. [16, 23] and in other areas of B.C. [24, 25]. In Kikomun Creek Provincial Park in the

Interior of B.C., Western Painted Turtles usually overwintered on or near the muddy bottom of lakes and ponds,

near to the shoreline at depths of about 1 m [24]. In the Williams Lake area in north-central British Columbia,

turtles overwintered at depths of less than 1 m, and occasionally less than 0.5 m, immediately adjacent to the

shore-line and buried in organic detritus amongst cattail roots [25]. Interestingly, the tagged individual was able

to locate this preferred habitat type in the Alaksen NWA wetland complex despite the habitat type’s rarity.

Emerging in this habitat type in the spring likely presents ample foraging and basking opportunities, and the

shallow stagnant water likely heats up faster than water in the other lagoons. This Western Painted Turtle had

to travel a great distance to reach this small wetland; however the distance reported for this individual (ca. 2.0

km) is not outside of the range of travel distances reported for Painted Turtles elsewhere [26]. Indeed recent

research suggests that freshwater turtles are actually very effective dispersers, regularly moving distances of up

to 1.3 km between wetlands, and as much as 3.3 km, both overland and through connecting water bodies [26].

Management Recommendations

Population Augmentation

There is considerable evidence that only a very small number of Western Painted Turtles inhabit this site, and

there is no evidence that successful reproduction is occurring. Thus this population is most likely inviable and

would require population augmentation to persist into the future. Due to the large size and extent of suitable

habitat in this area, as well as the protected status and length of time since this designation, this site has the

potential to support a large Western Painted Turtle population. Alaksen NWA could thus be a valuable asset for

potential re-introduction or supplementation of Western Painted Turtle populations on the south coast.



However, before reintroduction of this species is considered, it is highly recommended that the source of

agricultural pollution be addressed. Pollution may represent a significant impact on the suitability of this habitat

for maintaining a healthy population of Western Painted Turtles. Before any consideration of potential

augmentation of turtle populations at this site, some effort should be made to reduce pollutant levels in the

aquatic habitat. Although we did not find any abnormalities in any of the adult turtles that were captured,

exposure to pollutants are much more likely to have an effect on hatchling or juvenile turtles, and could further

explain the small, inviable Western Painted Turtle population observed at this site.

Invasive Species Removal

The base-line population data for the Red-eared Slider suggests that because they outnumber native turtles they

are likely competing for ideal basking, foraging, and nesting sites, and may be negatively affecting Western

Painted Turtles. Because it is now known through trapping efforts that Red-eared Sliders and Carp can be

successfully captured, this technique could be used as part of an invasive species management plan to remove

these species. Recent frog monitoring conducted at this site has also shown that Bullfrog and Green Frog

populations are thriving [11]. Because Alaksen NWA is a federally protected and federally managed site, invasive

species removal programs could be initiated and are recommended.

Habitat Enhancement

Although there is no evidence that Western Painted Turtles are successfully reproducing at Alaksen NWA, it

would still be recommended that enhancement take place at the potential nesting area in case there is a

possibility that turtles may reproduce successfully if nesting habitat was improved. The apparently preferred

location is the open area immediately east of the visitor parking lot, where there is an encroachment of invasive

vegetation (mostly blackberry) that is filling in the open area. This habitat could be greatly enhanced through

vegetation removal and addition of more suitable nesting substrate. The work would be neither time-consuming

nor expensive, and would result in significant enhancement of this nesting area. A closure of the visitor parking

lot would be recommended for the spring nesting season if and when Western Painted Turtles are confirmed to

nest at this site. In conjunction with this closure (mid-May) enhancement work could be undertaken.

Additional Research

Potential research into nesting habitat requirements, effect of habitat conditions on nest survival and hatchling

recruitment, and possible impacts of climate change on reproduction and recruitment of Western Painted

Turtles should be conducted at this site if Western Painted Turtles are confirmed to nest here. Although a

temperature/humidity data logger has already been installed, no nests have been confirmed. Monitoring for

Western Painted Turtle nesting should continue yearly and can be conducted by Canadian Wildlife Service

biologists. Currently, there is a lack of evidence that Red-eared Sliders successfully nest on the south coast, so

any identified Red-eared Slider nests should also be monitored.

Outreach and education

Because Alaksen NWA is open to the public, it would be advisable to install some interpretive signage at this site

to warn visitors of the possibility of nesting female turtles on land, as well as the importance of not releasing pet

Sliders into the wetlands. All interpretive signage should be approved and sponsored by the Canadian Wildlife

Service.



References

[1] The Western Painted Turtle Recovery Team. 2010. Draft Recovery Strategy for the Western Painted Turtle (Pacific Coast

Population), Chrysemys picta bellii, in British Columbia (March 2010). Original version prepared by Vanessa Kilburn for the

B.C. Ministry of Environment, Victoria, BC. 45 pp.

[2] Blood, D.A and M. Macartney. 1998. Painted Turtle. B.C. Min. Environ., Victoria, BC.

<http://wlapwww.gov.bc.ca/wld/documents/pturtle.pdf> [Accessed 7 Jul. 2009]

[3] Gregory, P.T. and R.W. Campbell. 1987. The Reptiles of British Columbia. B.C. Prov. Museum Handbook. Minist. of Prov.

Secret. and Gov. Serv. Victoria, BC. (from species description)

[4] Canadian Wildlife Service. 2001. Alaksen, British Columbia. Information sheet on Ramsar wetlands.

http://www.wetlands.org/RSIS/_COP9Directory/Directory/ris/4CA009en.pdf <Accessed on February 1st, 2010>

[5] Woudneh, M.B., Z. Ou, M. Sekela, T. Tuominen, and M. Gledhill. 2009. Pesticide Multiresidues in Waters of the Lower

Fraser Valley, British Columbia, Canada. Part I. Surface Water. J. Environ. Qual. 38(3):940-947.

[6] Wan, M.T., J. Kuo, and J. Pasternak. 2005. Residues of Endosulfan and Other Selected Organochlorine Pesticides in Farm

Areas of the Lower Fraser Valley, British Columbia, Canada. J. Environ. Qual. 34:1186-1193.

[7] C. Albert. Acting Protected Areas Coordinator, Canadian Wildlife Service. Delta, B.C. Email: [email protected]

[8] Crews, D., J.M. Bergeron, and J.A. McLachlan. 1995. The role of estrogen in turtle sex determination and the effect of

PCBs. Environ. Health Perspect. 103(Suppl 7):73-77.

[9] Shelby-Walker, J.A., C.K. Ward, and M.T. Mendonca. 2009. Reproductive parameters in female yellow-blotched map

turtles (Graptemys flavimaculata) from a historically contaminated site vs. a reference site. Comp. Biochem. Physiol. A.

154(3):401-408.

[10] Matson, C.W., G. Palatnikov, A. Islamzadeh, T.L. McDonald, R.L. Autenrieth, K.C. Donnelly and J.W. Bickham. 2005.

Chromosomal Damage in Two Species of Aquatic Turtles (Emys orbicularis and Mauremys caspica) Inhabiting Contaminated

Sites in Azerbaijan. Ecotoxicology, 14(5):513–525.

[11] Bell, B., J.R. Spotila and J. Congdon. 2006. High incidence of deformity in aquatic turtles in the John Heinz National

Wildlife Refuge. Environ. Pollut. 142:457-465.

[12] Maltby, F. L. 2000. Painted turtle nest site enhancement and monitoring, Red Devil Hill nest site at Revelstoke, BC.

Prepared for the Columbia Basin Fish and Wildlife Compensation Program, Nelson, BC.

[13] Cadi, A. and P. Joly. 2003. Competition for basking places between the endangered European pond turtle (Emys

orbicularis galloitalica) and the introduced Red-eared slider (Trachemys scripta elegans). Can. J. Zool. 81:1392-1398.

[14] Hays, D.W., K.R. McAllister, S.A. Richardson, and D.W. Stinson. 1999. Washington state recovery plan for the Western

Pond Turtle. W.A. Dept. Fish and Wildl. Olympia, WA.

[15] Committee on the Status of Endangered Wildlife in Canada (COSEWIC). 2006. COSEWIC assessment and status report

on the Western Painted Turtle Chrysemys picta bellii (Pacific Coast population, Intermountain- Rocky Mountain population

and Prairie/Western Boreal – Canadian Shield population) in Canada. Committee on the Status of Endangered Wildlife in

Canada. Ottawa, Ontario. vii + 40 pp. <www.sararegistry.gc.ca/status/status_e.cfm>.

[16] Ultsch, G.R. 2006. The ecology of overwintering among turtles: where turtles overwinter and its consequences. Biol.

Rev. 81:339-367.

[17] Sexton, O.T. 1959. Spatial and temporal movements of the painted turtle, Chrysemys picta marginata (Aggasiz). Ecol.

Monog. 29(2): 113-140.

[18] Blood, D.A and M. Macartney. 1998. Painted Turtle. B.C. Min. Environ., Victoria, BC.

<http://wlapwww.gov.bc.ca/wld/documents/pturtle.pdf> [Accessed 7 Jul. 2009]



[19] Steen, D.A. and J.P. Gibbs. 2004. Effects of roads on the structure of freshwater turtle populations. Conserv. Biol.

18(4):1143-1148.

[20] Garber, S.D. and Burger, J. 1995. A 20-yr study documenting the relationship between turtle decline and human

recreation. Ecol. Appl. 5(4):1151-1162.

[21] Marchand, M.N and J.A. Litvaitis. 2004. Effects of habitat features and landscape composition on the population

structure of a common aquatic turtle in a region undergoing rapid development. Conserv. Biol. 18(3):758-767.

[22] Bouchard, S.S and K.A. Bjorndal. 2006. Ontogenetic diet shifts and digestive constraints in the omnivorous freshwater

turtle Trachemys scripta. Physiol. and Biochem. Zool. 79(1):150-15.

[23] Mitchell, J.C. 1988. Population ecology and life histories of the freshwater turtles Chrysemys picta and Sternotherus

odoratus in an urban lake. Herp. Monog. 2: 40-61.

[24] St. Clair, R.C and P.T. Gregory. 1990. Factors affecting the northern range limit of painted turtles (Chrysemys picta):

winter acidosis or freezing? Copeia. 4:1083-1089.

[25] J. Steciw. Wildlife Biologist, B.C. Ministry of Environment. Williams Lake, BC. Email: [email protected]

[26] Bowne, D.R., M.A. Bowers, and J.E. Hines. 2006. Connectivity in an agricultural landscape as reflected by interpond

movements of a freshwater turtle. Conserv. Biol. 20(3) 780-791.

Appendix A:

ata Collection Forms Surveying and Monitoring Protocols and Associated D

) Chrysemys picta bellii Western Painted Turtle (

h @ cloaca and plastron (cm) Total tail length, tail length to cloaca, tail widt

Right and left front middle claws (cm)

Carapace width and length (cm)

Plastron width and length (cm)

Weight (kg)

Height (cm)

Data Collection Form 3): plastron, head and left/right front feet claws (see

pace, ng characteristics and photos are taken of the caraOnce captured, turtles are measured for the followi

once every two to three days.

least keep turtles interested. Bait should be changed at types can be alternated within a trapping event to

d bait ultiple bait types can be used in a single trap, anyellow croakers, or cans/packets of wet cat food. M

such as ed sardines, or other smelly frozen fish varieties capture turtles. Hoop traps can be baited with cann

well as opportunistic hand-capture can be used to techniques are employed. Hoop and basking traps as

cument population demographics, mark/recapture To more accurately assess relative abundance and do

). estimate of abundanceof the population and provide only a very relative

kely to detect only a portion even on optimal survey days, basking surveys are liNote:population abundance. (

ve ls observed basking to give an indication of relatiidentified to determine maximum number of individua

n be concentrated in areas where turtles have been basking surveys on optimal days by boat or shore ca

population assessments can be conducted. Intensive Once turtle presence has been confirmed at a site,

Population Assessments

General Monitoring Protocols

observed or not.

es are e visit regardless of whether Western Painted TurtlHabitat assessments should be made on the first sit

n Form 2). eats present at the survey site (see Data Collectiodetermine habitat characteristics and potential thr

e to ection Form 1) and habitat assessments are also madObservations of turtles are recorded (see Data Coll

non-detections.

dence in h location on optimal survey days to increase confiIdeally at least three surveys are conducted at eac

. oon in late March-early May and again in September)temperatures are lower than air temperature (near n

days when turtles are active and average water The optimal time to survey for turtles is on sunny

ng turtles. e south facing have a higher likelihood of attractilarge enough for a turtle to rest its body) that ar

mb and g., not too far out of the water for turtles to cliAreas with woody debris appropriate for basking (e.

ee. presence prior to potentially causing turtles to flbody) and look from a distance to determine turtle

ater y from shore (> 50 m away or in the middle of the wWhen surveying by boat, care is taken to paddle awa

by boat, a spotting scope may be used.

lt to access ult to see basking locations from shore and difficufrom shore or by boat. For areas where it is diffic

10 am – 3 pm) and can be conducted using binocularsSurveying for WPT occurs during peak basking time (

Basking Surveys

General Survey Protocols

•

•

•

•

•

•

•

•

o o o o o o

females of the population.

y for rtles to add to population assessment data, but onlan alternative time to mark/recapture individual tu

so be ored to determine number of nests laid. This can alobserved nesting or attempting to nest can be monit

en sting season (May-July), areas that turtles have beIn the evening (between 5:00 and 9:00 pm) during ne

ithin 100 m from the occupied water body. ground cover vegetation and limited canopy cover, w

h limited south-facing on gently sloping or flat terrain, witareas of sandy, rocky or clayey substrate that are

mely, tified based on certain habitat characteristics; naPotential nesting areas to be monitored can be iden

is confident the nesting attempt is complete.

erver ever, turtles should only be disturbed once the obsopportunity to mark any un-notched individuals. How

urtle activities. Nesting surveys can be used as anpopulation and provide more information on marked t

ar turtle entify key threats that may be affecting a particulthis aspect of a turtle population also helps to id

itoring ons to determine fecundity and nesting success. MonNesting surveys are used to monitor turtle populati

Nest Site Monitoring

. Notching on a turtle carapace Figure 1

Individual 2

n Example: Right scute marking indicates Individual number – I

39 Left scute indicates site number – In example: Lake

done with a triangular file.

otching is individual number on the right (Figure 1). Shell n(Figure 1). Site number is notched on left side and

isation e thickest part of the shell with the most vascularpurposes, scutes 4, 5 and 6 are skipped to avoid th

marking top of the carapace behind the head. However, for carapace , excluding one small central scute at the

e of their artitions on the edge of their shell) on either sidcarapace (Figure 1). Most turtles have 12 scutes (p

of their turtles are marked by shell-notching on both sides To identify individual turtles for mark-recapture,

protocol.

a detailed s and adults are easily sampled. See Appendix B forup to 10 turtles per location. Only larger juvenile

on size; and the number of samples is based on the populatiBlood samples are also collected at every location,

are also noted. Any special identifying features, such as scarring,

plastron)

al distance of their tail to their cloaca from the smaller, have longer claws, and a longer proportion

d overall appearance (e.g., males are generally Sex and age are determined based on measurements ano

o

•

•

•

•

•

(e.g., coming closer to the water surface).

e turtle signal strength are not just due to movement of thestimate the actual location, and that any jumps in

y can ource from at least 2 locations until confident thesignal. Observers obtain directions to the signal s

istent n picking the direction of the strongest, most consthe signal drops and where it is the strongest, the

g where ated by moving the antenna back and forth, detectinWhen the turtle is not visible, locations are estim

the signal. ut-in location for the boat is near the location oflocations will likely bias turtle movement if the p

se re when its location is close. Using a boat for theObservers may estimate the turtle position from sho

n. ing should be conducted even with visual observatioapplicable; although a test of transmitter function

on when d and used in replace of location of signal detectiVisual observations during the day are also recorde

) and 3-element YAGI antenna. (2006) using a portable receiver (Wildlife Elements

stent signal) based on methods used by Bowne et al.boat (marking GPS location at strongest, most consi

e or by osition either by estimating the location from shorA transmitted turtle is located via homing to its p

rapace. the bolts are located on the top of the turtles’ ca

e so that be threaded from the bottom of the turtle’s carapacplace with stainless steel nuts. The screws should

which are held in smitter is then affixed with stainless steel bolts held electric drill and a small drill bit. The tran

th a hand-er holes, about 2 cm from the edge of the shell, withrough the scutes in the position of the transmitt

led nted in Figure 1). To do this, holes are first drilscutes numbered 6 and 7 in the marking scheme prese

and 10, or via bolts to two posterior scutes (usually scutes 9Radio-transmitters can be affixed to adult turtles

Radio-Tracking

Radio-Tracking, Movement and Habitat Use

early as September of the year the nest was laid. June the year following laying, but could occur as

h-tchling emergence, which generally occurs from Marcby humans. Exclosures should be removed prior to ha

targeted the exclosure is completely buried and will not be Samson et al. (2007), and can be installed so that

s in ing plastic hardware cloth and heavy metal stakes afrom predators. A predator exclosure can be made us

ect eggs ator exclosures may be installed over nests to protIf predation is a concern at the nesting site, pred

(ca. 10 cm).

t the nest at approximately the same depth as the nesthem from the elements and buried about 25 cm from

rotect s. Data loggers should be placed in nylon mesh to pthe seasons for long-term climate change assessment

ughout he nesting sites to monitor temperature trends throTemperature data loggers can also be installed at t

. where the distance between the two markers is known

B), istances to the nest off of permanent markers (A & may be located via triangulation by measuring two d

a nest te for the nesting area, the exact UTM position of marking a nest with a visible marker is inappropria

. If ld make turtle nests a target for human disturbancethe markers are not obvious to the public which cou

re that a public nesting area, care should be taken to ensurecruitment can be determined. If marking nests in

g through to the next season is possible and hatchlinWhen possible, nests are marked so that monitoring

llection Form 4). as the date, time, and air temperature (see Data Co

e, as well k the female to complete nesting, the substrate typlaid if the attempt was successful, how long it too

were sting attempt was successful or not, how many eggs Observations are recorded, including whether the ne

r to nest hole digging starts. approaching the turtle or making any movements prio

avoid ve to disturbance and thus care should be taken to Before a female starts to nest, she is very sensiti

10 m). tance to avoid disturbing nesting females (at leastNest-site monitoring should be done from a safe dis•

•

•

•

•

•

•

•

•

to get signals.

ore locations ion of the turtle and the accessibility of ideal shincreasing error of location depending on the posit

luencing turtle movement (due to boat presence) andThe method of choice may be a trade-off between inf

GPS, and compass would be needed.

ver unit, re 2). However, for this method an additional receiangles would be the estimated turtle location (Figu

f the . The point of intersection between the two lines ovia GPS) and the angle of strongest signal strength

arked MapSource by inputting each observer’s location (mturtle location. Locations would be estimated using

angle as possible with the es to create as close to a 90minimize error, observers need to position themselv

t [2]. To ure 2) on land while being in constant radio contaccompass) from 2 different locations (A and B in Fig

using a ection of the radio signal and recording an angle (people simultaneously determining the strongest dir

to determine the turtle location; this requires 2 triangulatingAn alternative tracking method would consist of

’ the location (as described below). ‘triangulategle person can be sufficient to derive an accurate location, a sin

ill not r other GPS mapping software). In cases when that wthe turtle later via plotting on a MapSource map (o

dinates of ation in order to accurately determine the GPS coordistance to the turtle and a description of the loc

coordinates are recorded along with an estimated When turtle locations are estimated from shore, GPS

overwintering site. weather, especially during the winter months at the

nclement g during evening tracking sessions) and/or during inightly locations (due to safety concerns of boatin

btaining hod of locating turtles may also be utilized when oaccessible shore but an adaptation of the shore met

n appropriate for daily locations that are far from aThe method of using a boat to acquire locations is •

•

•

o

•

ion using radio-telemetry. . Triangulation method for determining turtle locat Figure 2

90 degree difference is

ideal

eaf litter, on; substrate can be classified as organic (e.g., laccomplished by dipping a paddle at the plot locati

sily en to look at substrate composition, and is most eaAn aquatic sediment/vegetation sample should be tak

bmergents.typically clear of vegetation and woody emergent/su

m edge on either side of the water body that is Open water habitat (OP), the area outside of the 10

itat but can also exist in open water. Basking habitat (BA), often located within Edge Hab

fall from the riparian habitat.

woody emergents/submergents because of dead vegetation (including trees) and generally has more

m of shore which can be shaded by riparian Edge Habitat (ED), referring to the area within 10

Dalgarn (2009)]:

we and ed as follows [partially based on definitions by RoGeneral habitat descriptions to be used are describ

species visibly present in immediate area

ts in 2 m radius, other fish, wildlife, plant of canopy), degree of woody emergents and submergen

clear ge, open, basking), degree of cover (i.e., shaded, General habitat description: habitat type (i.e., ed

Water depth (using pre-marked rope with weight)

Conductivity

pH

sking season only)Nearest basking location temperature (during the ba

Bottom water temperature

Water column temperature (mid-way)

Surface water temperature

Air temperature

of each location (see Data Collection Form 6):

racking d be measured/recorded as soon as possible to the tThe following aquatic habitat characteristics shoul

weekly or monthly tracking events.

n on d during daily tracking events, or at every locatio>10m apart (average error of locations) if conducte

hat are t plots can be conducted at all tracked locations tTo compare habitat use daily and seasonally, habita

Habitat Use

movements (daily and seasonally).

e the data set for a more comprehensive look at turtlTrapping and recapture locations can also added to

habitat. [mid-March] are obtained to identify overwintering

ring /winter [October-November] and late winter/early spAdditional radio-tracking locations during the fall

Collection Form 5).

see Data h observations of turtle activity when applicable (season [July-September]. Locations are recorded wit

active urs (7 locations per day = 28 locations) during thedays (8am – 8pm) with locations obtained every 2 ho

our study ve radio-tracking should be conducted on at least fbased on Crocker et al. (2000) can be used. Intensi

dy design patterns during its active season, an intensive stuTo identify the turtle’s daily and weekly movement

Movement Patterns

•

•

•

•

•

o

o

o

o

o

o

o

o

o

•

o

o

o

•

Random out the available aquatic habitat for comparison.locations (excluding overlapping locations) through

tracked on of random points roughly equal in number to theHabitat plots should also be conducted at a selecti

) based on Marchand and Litvaitis (2004).woody debris) and inorganic (e.g., silt, sand, clay

•

http://www.spatialecology.com/htools/rndpnts.php

sis Tools for ArcGIS (source: plot locations can be generated using Hauth’s Analy

osition tegorized via habitat categories (or landscape compIf turtles are tracked on land, locations may be ca

)

•

References

Weighted rope with depths pre-marked

Glass thermometer

Temp/Cond/pH meter

Notebook and pencil

GPS unit (x 2 for triangulation)

Receiver and antenna (x 2 for triangulation)

Chest waders

Binoculars

Kayak, paddle, and life jacket

Equipment needed

sed on studies by Marchand and Litvaitis (2004).categories) already described in the report body ba

o

o

o

o

o

o

o

o

o

by interpond nectivity in an agricultural landscape as reflectedBowne, D.R., M.A. Bowers, and J.E. Hines. 2006. Con

) in Rhode Island. Can. J. Zool. 78:936–942 Chrysemys picta pictapainted turtles (

of eastern ackson. 2000. Overwintering behavior and physiologyC.E. Crocker, R.A. Feldman, G.R. Ultsch, and D.C. J

(3):780-791. movements of a freshwater turtle. Conserv. Biol. 20

opulation habitat features and landscape composition on the pMarchand, M.N and J.A. Litvaitis. 2004. Effects of

43:926-935.ling error in radio triangulation. J. Wildl. Mgmt.Springer, J. T. 1979. Some sources of bias and samp

er turtles. Chel. Conserv. Biol. 6(2):255-259. morphometric data from small-sized eggs of freshwat

fecundity and xcavation is a non-deleterious method for obtainingSamson, J., E. J. Hughes, and R. J. Brooks. 2007. E

. 1:85-92 Painted Turtles (Chrysemys picta marginata). Copeia

ion in Midland nd Microhabitat Use on Diel Body Temperature VariatRowe, J.W. and S.J. Dalgran. 2009. Effects of Sex a

8-767. dergoing rapid development. Conserv. Biol. 18(3):75structure of a common aquatic turtle in a region un

If found please contact HAT 250.995.2428 or email [email protected]

Data Collection Form 1: Turtle Basking Survey Form

Modified: May 21, 2010

Survey Information

Location name:

Observer(s):

Date (dd/mm/yr):

Time: Start: End:

Person-Search effort: hours =

Foot Canoe KSurvey method: ayak Powerboat Other:

Temperature (ºC): (at 10 cm depth in ca. ½ m deep water)

Air : Water:

Calm LightWind speed:

Moderate Strong

Binoculars Telescope Viewing equip.:

Precipitation: None Fog Light Moderate Hard Other

%Cloud cover (%):

Clear <50 >50 Overcast

estimate or

Seen Heard No Bullfrogs seen:

Species Observation(s) Information

Map reference

Species1 # seen Activity2 Microhabitat3 If possible Sex M/F/U Size class4

Notes on observations

(mark locations with letters/numbers/WPT #s): Site sketch or attach map

l = <6cm Estimated size: Large >9cm; Medium = 6 – 9cm; Smalher (describe in comments) K = rock; RHIZ = rhizome mat; SHORE - shoreline; Ot LOG= on log; Water = in water; Land = on land; ROC

alking on land nesting , digging on land; TS = swimming; TW = w BA = basking; CO = courtship; FD = feeding; NE= RSC) or other (identify or take a picture) Western Painted Turtle (CHPI); Red-eared Slider (T

1

2

3

4

.995.2428 or email If this spreadsheet is found please contact HAT 250 [email protected] Modified: May 22, 2010

nt Data Collection Form 2: Habitat and Threat Assessme

Date: (+ camera owner’s initials):Photo #’s Location Name:

Observer(s):

(Use NAD83 datum) Northing: UTM: Zone U Easting:

Type of Ownership: Private Park Crown Federal Other: Landowner contact info: Landscape context: Urban Rural Agricultural Backcountry Other:

SITE DESCRIPTION AND ASSESSMENT (in large water bodies only for surveyed areas) :

Bottom substrateLake Creek River Other: : Marsh Slough Natural-pond Human-made pond Wetland Type

:

(10 m zone from shoreline)Aquatic vegetation cover

er: Fines/muck Detritus/organics Sand Gravel Oth

: _____% emergent; _____% floating; _____% submergent

Dominant aquatic plant species:

Shoreline zone cover (10 m inland from shoreline): ____%forest; ___%shrub; __ %grass/herb; __%developed

(40 m beyond shoreline zone):Upland cover

___%other (describe): _

____%forest; ____%shrub; ____%grass/herb; ____%developed

___%other (describe): _

Describe upland habitat:

Basking

site type: Logs Rhizome mats Rocks Shrub overhang Shoreline/banks other:

Assess basking opportunities: Nesting habitat: Confirmed; Potential; None identified

Describe nesting habitat/assess opportunities (indicate on map):

Assess habitat suitability (loss, condition, movement barriers etc.):

ON-SITE THREAT ASSESSMENT:

(within 50 m from water’s edge or as visible): B) Threats to aquatic habitat and travel routes

A) Threats to nesting habitat (describe):

mentRoads - paved H M L n/a Housing/industrial develop 1

Roads - unpaved H M L n/a Urban activities H M L n/a

2

Introduced species YesPets H M L n/a Agriculture H M L n/a

H M L n/a Recreation - non motorized H M L n/a Grazing H M L n/a Recreation - motorized H M L n/a Logging

H M L n/a

3 No

: Name of introduced species

n/a Water use /control H M L n/a

C) Comments on threats:

ccurately Yes-Known to be present; risk cannot be assessed aassociated with housing/urban developments Landscaping, gardening, or other human activities

In progress, planned or potential

1

2 3

Recorder Name:

ation Assessment Information Data Collection Form 3: Turtle Mark-Recapture Popul

Date:

Weather:

Recorder Name: Date:

Weather:

Location Description (in detail): Circumstances of Capture:

WYPT ID:

GPS – Easting:

GPS – Northing:

Turtle Species: Age:

Sex: Gravid?

Carapace length (mm): Carapace width (mm):

Plastron length (mm): Plastron width (mm):

Height (mm): Weight (mm):

Tail total length (mm): Tail length to cloaca (mm):

: Tail width at shell (mm): Tail width at cloaca (mm)

gth (mm): Left middle claw length (mm): Right middle claw len

Injuries or distinctive features:

Location Description (in detail): Circumstances of Capture:

WYPT ID:

GPS – Easting:

GPS – Northing:

Turtle Species: Age:

Sex: Gravid?

Carapace length (mm): Carapace width (mm):

Plastron length (mm): Plastron width (mm):

Height (mm): Weight (mm):

Tail total length (mm): Tail length to cloaca (mm):

: Tail width at shell (mm): Tail width at cloaca (mm)

gth (mm): Left middle claw length (mm): Right middle claw len

Injuries or distinctive features:

TURTLE NESTING SURVEY

Data Collection Form 4: Turtle Nesting Survey

TURTLE NESTING SURVEY

Recorder Name:

Recorder Name:

Date:

Date:

Weather:

Weather:

land? spotted on Turtle

Time End

Time Start

Location Survey

on land?spotted Turtle

Time End

Time Start

Location Survey

TURTLE NESTING ATTEMPT OBSERVATIONS

TURTLE NESTING ATTEMPT OBSERVATIONS

Location Description (in detail):

Location Description (in detail):

GPS Location/coordinates:

GPS Location/coordinates:

Turtle Species: Already Marked?

Turtle Species: Already Marked?

Notch Code or Radio Tag # (new or existing):

Notch Code or Radio Tag # (new or existing):

Obs. Start Time: Obs. End Time:

Obs. Start Time: Obs. End Time:

Substrate:

Substrate:

Distance from Water: Attempt Successful?

Distance from Water: Attempt Successful?

Photos: # of Eggs:

Photos: # of Eggs:

For mapping and nest re-locating purposes

For mapping and nest re-locating purposes

installed? Predator exclosure

Nest I.D.

installed? Predator exclosure

Nest I.D.

Distance from A Distance from B

Distance from A Distance from B

Describe Turtle Activity/Comments:

Describe Turtle Activity/Comments:

: by boatI (1) or (2): Triangulation - 1 or 2 people, and BO*Track type: VIS: Visual, SH: by shore (1 person), TR

CommentsStrength

Signal Triang A Triang B Actual Triang A Triang B

type*Track

Point # Time FreqSessionTrack

Bearing (degrees)Location NAD 93 10 U

Weather:Date: Observer:

Data Collection Form 5: Turtle Radio-Tracking

Track/R

type*Plot

General habitat characteristicsTemperature (

Data Collection Form 6: Turtle Habitat Use Plots

o)

adom Pt

: TR: tracking location, RA: random location*Plot type

Comments(Y/N)

Pic vis****Water

Veg***Submerg Emerg /

**Submerg*Emerg / Woody

type**habitat Aquatic

(cm)Depth

Conduct PhbaskNear

WaterBottom

WaterColumn

WaterSurface

Date Time AirLocat

**Aquatic habitat type ear of cover ; includes Degree of Cover: SH: Shady (%) or CE: Cllassifications (Rowe et al. 2009)), and BA: Basking: OP: Open water, ED: Edge (based on microhabitat c

***Emerg / Submergs itvatis 2004)) within 2 mt (based on basking site categories (Marchand and L: 0 - sparse, 1- scattered, 2 - common, 3 - abundan****Water vis - clear: 0 - completely opaque, 1 - 50% transparent, and 2

by Elinor Hughes

nvironment Prepared for the British Columbia Ministry of the E

) blood and tissue for genetic analysis. Chrysemys pictaturtle (

f painted Appendix B: Protocol for the sampling and storage o

............... 19...................................................Appendix 2: Suggested suppliers ...................

18s ................................................ Appendix 1: Cost comparison of blood storage method

............................. 17...................................................References ........................................

................. 16 ...................................................Possible laboratories ...................................................... 16...................................................Testing guidelines ................................

....................... 15 ...................................................Method ..................................................................... 15...................................................Tail clipping .....................................

....................... 14 ...................................................Method .................................................................... 14...................................................Toe clipping ......................................

.......................... 13...................................................Materials .........................................

................ 13...................................................Storage of tissue samples .........................

............... 13...................................................Toe clipping vs. tail clipping ....................

......................... 13...................................................Tissue Sampling ...................................

urtle tail ............................ 10 Some points to remember when drawing blood from a t.................... 9 ...................................................Drawing blood .....................................

................. 9 ...................................................Preparation of materials ...................................... 9...................................................Turtle blood sampling method ......................

................. 8 ...................................................Making cell lysis buffer ...................................................... 8...................................................Materials .........................................

...... 7...................................................Comparison of blood storage methods ...............

................. 7...................................................Storage of blood samples ..........................

............................ 7...................................................Blood sampling ....................................

........... 6...................................................A note on genetic variability in turtles ..........

............................... 5...................................................Tissue ............................................

............................... 5...................................................Blood .............................................

.......................... 5...................................................Sampling protocol .................................

............................ 3...................................................Table of Figures ..................................

2

............... 12 ...................................................relation to the yellow stripe. ....................oaca and to the end of the tail and in insertion of the needle, both in relation to the cl

pace) and the location for the turtles (female cloacas are much closer to the caraimate location of the cloaca in male Figure 2 Dorsal view of a turtle showing the approxgle from vertical. ............................ 11 and needle inserted into turtle tail at a slight an

of caudal artery within caudal vertebra Figure 1 Turtle tail cross-section showing location

Table of Figures

3

possible genetic markers and testing laboratories. c testing guidelines, including methods and sample storage requirements; and genetiirements; tissue sampling, including sampling, including methods and sample storage requ

cs include: sampling protocol; blood turtles for the purposes of genetic analysis. Topion of samples from painted The following comprises a protocol for the collecti

eatened species. management as part of a recovery strategy for a thrtion) and 5) the potential for genetic recent (e.g. a new road) or historical (e.g. glacia) the effect of an isolation event, either presence of inbreeding and its effect on fitness, 4

ith a loss in specific phenotypes, 3) the example whether a loss of diversity is associated w diversity in small populations, for phylogeography, 2) the effects of a loss of geneticships among sister species and population or species, including taxonomic relation1) the evolutionary history of a Genetics may be used to study questions regarding: te without including genetic analysis. of within-or among-population diversity is incomple

, many may suggest that an investigation useful and achievable in the recent years. In factncreasingly more important, The use of genetics to study diversity has become i

4

Blood

Sampling protocol

5

Tissue

turtles caught. that blood be drawn from all adult the sex ratio is unknown, it is highly recommended

caught. If either the population size or used will depend on the how frequently turtles are caught each day. The sampling regime day or draw blood from the first turtle of each sex9:00 am, 12:00 pm and 3:00 pm each blood from turtles of each sex caught at (or near)

female turtle caught, draw nd male and every 2ndare as follows: draw blood from every 2f catch intervals that may be used random sample and is much simpler. Some examples o

pecific intervals produces an equally instructions regarding their use, but sampling at s be found on the Internet with specific catch interval. Random numbers tables mayble or by sampling turtles at a determined either by consulting a random numbers ta

ratio. The sampling regime may be sex ratios, the sample group should reflect the sex sampled. In populations with uneven sampled or the absolute number of individuals to be

the proportion of the population to be before you run out of catchable turtles. Determineblood from the required number and that the sampling regime allows you to collect

n, ensure that the sample is random may be used. When taking a sample of the populatioconcern, then a sample of the population populations are small. If time or resources are a

es in a population, especially if Ideally, blood should be drawn from all adult turtl

the individual hatchlings. randomness of the sample applies to the nests, not als within a single nest, and so the For nests, tissue should be taken from all individuabove for adult turtles may be used. nests, a similar sampling regime to that described y be used. For both juveniles and or 50 juveniles, then a sample of the population ma

sampled. If there are more than 20 nests juveniles, then all hatchlings/juveniles should be , e.g. fewer than 20 nests or 50 If few nests or juveniles are found in a population

performed. uals if a paternity analysis is to be samples should only be taken from non-adult individ

tion. Furthermore, we recommend that negative effects on the individual or on the populaor juvenile turtles to limit possible tissue samples should only be taken from hatchling

we conservatively recommend that the genetic diversity of a population. Therefore, may prove to be irrelevant to studies of is very slight. As a result, their genetic makeup will become a reproductively active adult populations, the likelihood that a juvenile turtle

g and juvenile turtles in natural Because of the very low survivorship of hatchliny active turtles (pers obs.) been observed on numerous healthy and reproductivel

l or complete loss of tails or limbs have Serious adult injuries, however, involving a partia of turtles has not been studied. The effect of tissue removal on long-term survival

A note on genetic variability in turtles

6

0 years ago. among populations isolated from one another even 10be observable genetic differences slower rate into consideration, e.g. there may not

researchers are advised to take the researching many of the among-population questions, conversion value. Hence, in These numbers are only included as a caution, not ato a “conventional” mt-DNA rate. DNA) differentiation in turtle species as compared

ower rate of mitochondrial DNA (mt-other species. Avise et al (1992) found a 2-14x sled to change as quickly as in Genetic variability in turtles should not be expect

Storage of blood samples

Blood sampling

7

Comparison of blood storage methods

hat the samples remain desiccated. have dried, however, care must be taken to ensure t at room temperature, once the samples stored on cell lysis cards may be kept indefinitely

care must be taken. Blood samples but freeze-thaw cycles may fragment DNA strands so for long-term storage, C). Samples may be stored at subzero temperatures refrigerator (4

but must then be kept in a may be held for up to one week at room temperature,lood samples stored in a cell lysis buffer analysis: cell lysis buffer or cell lysis cards. B

lood samples for future DNA There are two field-convenient methods of storing b

°

cards for painted turtle blood storage. analysis, we would recommend the use of cell lysis e detail. Based on this cost-benefit section and the laboratory analysis section for mored laboratory costs. See the materials materials for drawing blood, and both have associatonally, both methods require method are included in the cost comparison. Additi

y those materials that are unique to each suggests suppliers for the required materials. Onl two methods and Appendix 2 Appendix 1 outlines the cost comparison between the

1) Slightly more expensive per sample. Cons

very fast (30 minutes to usable DNA). 3) The extraction technique is fairly simple and isough the mail. 2) Samples may easily be transported, including thr

tely. 1) Samples may be kept at room temperature indefiniPros

Cell lysis cards:

blood-taking. uld be refrigerated with 24 hours of acceptable (up to a week), but ideally, samples short durations at room temperature are 3) The samples must be kept in a refrigerator. Sho

ampling many individuals 2) The tubes are bulky to carry and cumbersome if sd then autoclaved. chemicals, but must be adjusted to a specific pH an

It requires common laboratory 1) Cell lysis buffer must be made in a laboratory. Cons

ults. generally needed to produce enough DNA for good resnly a very small amount of blood is however, since turtle blood cells contain nuclei, o

may result in a higher DNA yield, 2) The extraction process may be more efficient andosed to ~$4.20/sample). 1) Slightly cheaper per sample (~$2.80/sample as opp

Pros

Cell lysis buffer:

Materials

esearchers. materials be placed by a manager and dispensed to rurther recommend that orders for are only obtainable in large quantities, we would f

below. Because many of the items Regardless, we include descriptions of both methods

8

storage has not been tested. ever their effectiveness in turtle blood Note: lysis buffers are available commercially, how

samples. er and will affect the quality of the evaporation of liquid will alter the pH of the buffthe caps are tightly closed as any indefinitely. Care should be taken to ensure that

be kept at room temperature be prepared well in advance of blood taking and mayady to draw blood. Multiple tubes may close cap tightly and place in storage box until re

into each 1.5ml microcentrifuge tube, Once the buffer is finished, dispense 1ml of bufferfer should be autoclaved before use. workers will find the recipe easy to read. The buftories. Additionally, most laboratory chemicals that are readily available in many labora

-lauroylsarcosine, pH 8.0. These are all common NaCl, 0.01 M sodium EDTA and 10% s as follows: 0.01 M Tris, 0.01 M specific pH. The recipe for Queen’s lysis buffer i

and soaps that is standardized to a al., 1991). A buffer is simply a solution of salts Queen’s lysis buffer (Seutin et If using buffer to store turtle blood, we recommend

Making cell lysis buffer

ot available. 1cc syringes attached might be used if others are nh skin. That said, 29 gauge needles with may better resist bending against the turtles’ toug

les are recommended because they smaller needle diameter). The larger diameter need or smaller (larger gauge number = needles already attached and are generally 29 gauge

most drug stores, however, they have Note: 1.0ml (1cc) volume syringes are available at

rage boxes (for tubes) Storage bags and desiccant (for lysis cards) or stosis buffer in microcentrifuge tubes) Blood storage medium (either cell lysis cards or ly

Small bucket (optional) Fine permanent black marker Dishtowels Alcohol wipes (purchased at drug store) Sharps disposal container

ml syringe only 1.027 gauge bevel-tip needles

zed materials) (See Appendix 2 for suggested suppliers of speciali

n

Turtle blood sampling method

9

the needle to face the bevel in different have 0.1ml of blood. If there is no blood, rotate patiently draw on the plunger until you begin to flow into the needle. If there is blood,

n the plunger; you should see blood withdraw the needle very slightly and gently draw o. Once your hand is positioned, needle firmly in the tail--there is no need to rush

unger. Be confident and keep the 3) Reposition your hand so that you can draw the plbetween two vertebrae.

means that you have found a space until you feel the needle go in 1/8” to 1/4” . Thisy closer to the carapace. Repeat this immediately remove the needle and insert it slightl

Figures 1 and 2). If you hit bone needle will be at a slight angle from vertical, see Aim for the centre line of the tail (the the end of the tail just inside the yellow stripe.

the tail about one-finger’s width from 2) Gently insert the needle into the dorsal side ofhand.

and forefinger of your non-dominant facing up. Hold the tail firmly between the thumb e that the bevel of the needle is 1) Take the syringe in your dominant hand and ensur

Drawing blood

cohol wipe. another dishtowel and swab it thoroughly with an alwith a wet dishtowel, dry it with firmly with your non-dominant hand, clean the tail lean over as much. Grip the tail small bucket under your feet so you do not need to r dominant hand. If desired, place a your knees, tail up and carapace facing towards youound, hold the turtle firmly between being pinched. Seated on a chair/stump/rock/the gr

head are retracted into the body without secure the hind feet and ensure that all limbs and wel leaving the tail exposed. Be sure to 5) Prepare the turtle. Wrap the turtle in a dishto

t do not remove from packages. 4) Take two alcohol wipes and open the packages, buut a short way. package. Prime the syringe plunger by pulling it o

but don’ t remove it from the syringe package and loosen the cap from the needle,ringes with needles attached, open the plunger by pulling it out a short way. If using sy

eedle package. Prime the syringe cap from the needle, but don’ t remove it from the nfirmly onto the needle. Loosen the syringe and remove from package. Seat the syringe

e top of the package. Take one 3) Prepare the needle. Take one needle and open thof the tube.

arker. Write the turtle’s ID on the cap the side of a tube with a fine permanent felt tip mjuvenile, and the date in the frosted area on If using lysis buffer, write the turtle’s ID, sex/

provided by the manufacturer. familiarized yourself with the lysis card protocol age bag. Be sure that you have marker. Place one bag of desiccant inside the stortorage bag with a fine permanent felt tip sex/juvenile, and the date on the card and on the s

cell lysis card, write the turtle’s ID, 2) Prepare the cell lysis card or tube. If using asampling, difficulties with sampling).

le tail condition, injuries caused by regarding the blood-taking procedure (e.g. pre-samp, researcher’s name, comments turtle ID, sex/juvenile, population, date of sampleing. Column headings should include: 1) Create a data sheet to keep track of blood sampl

Preparation of materials

sample must be taken. done on those individuals, a tissue blood from hatchlings. If genetic testing is to bejuvenile turtles and impossible to draw 7) It is very difficult to draw blood from smaller

e and use a new needle if necessary. become bent. Be sure to check the tip of the needlt the needle repeatedly, the tip may 6) Turtle skin is very tough. If you need to inser

blood-taking technique. this if you are confident in your between the cloaca and the carapace. Only attempt

attempt to draw blood from the tail 5) If the turtle has a severe tail injury, you may ae. will be trying to insert the needle between vertebr

within the vertebral column. You 4) You are aiming for the caudal artery, which runs cloaca into the turtle’s body. that you will hit any vital nerves leading from the to the cloaca. This reduces the chance 3) Be sure that you are inserting the needle distal

turtle will kick at the syringe. bs should also be well wrapped, as the the researcher might not want bitten. The hind lim

head will be very near to body parts that turtles are small, they have fierce bites, and the secured in the towel. While painted 2) Ensure that the turtle’s head and limbs are well

to receive the sample) ollection medium labelled and ready seated on syringe and primed, alcohol wipes open, c

erials at hand and ready to use (needle on the tip. Be sure that you have all required mat, especially the location of the bevel 1) Familiarize yourself with the needle and syringe

urtle tail Some points to remember when drawing blood from a t

utes. injection site should be checked again after 30 minor swelling after 5 minutes, and the should be applied if there is any sign of bleeding nutes for bleeding or swelling. Bactine 6) Check the injection site after 1 minute and 5 mi

within about 30 seconds to1 minute. the turtle. Any bleeding should stop 5) Swab the tail with the alcohol wipe and release

container. x. Dispose of the needle in a sharps blood with the buffer. Put the tube back in the bo

ghtly closed and invert the tube to mix the the blood into the tube. Ensure that the cap is tit. If using tubes with buffer, dispense minutes and then place in storage bag with desiccan

Allow sample to air-dry for 30 the blood does not go beyond the circle indicated. ollection area on the card. Be sure that the blood lysis cards dispense the blood onto the caw blood to the 0.2ml mark. If using 4) Draw 0.1ml of blood. If the blood is aerated dr

stop trying and come back to that turtle later. vertebrae. If the turtle is very resistant, will find it difficult to insert the needle between

s. If the turtle is pulling in its tail, you blood. Some turtles are harder to bleed than othern another spot. Keep trying until you get directions, or remove the needle and re-insert it i

10

11

gle from vertical. and needle inserted into turtle tail at a slight an of caudal artery within caudal vertebra Figure 1 Turtle tail cross-section showing location

12

relation to the yellow stripe. oaca and to the end of the tail and in insertion of the needle, both in relation to the cl

pace) and the location for the turtles (female cloacas are much closer to the caraimate location of the cloaca in male Figure 2 Dorsal view of a turtle showing the approx

Toe clipping vs. tail clippingTissue Sampling

13

Storage of tissue samples

digit. cher’s comfort level with removing a is used for hatchling turtles depends on the resear

sue is all that is required. Which method samples are stored correctly, a single piece of tislip than by tail-clip. Nonetheless, if a larger amount of tissue may be collected by toe-c

ail sample may be taken. Hence (ASIH 2004; we recommend fewer), whereas only one tcent toes on each foot may be sampled hatchlings or juvenile turtles. Up to two non-adja

n, while tail clipping may be used on from the nest or caught in their first active seasoles, i.e., those sampled directly Toe clipping should only be used on hatchling turt

Materials

sampling. the buffer described in the above section on blood the protocol for making and dispensing indefinitely. To use Queen’s lysis buffer, follow

C samples may be refrigerated at 4temperature for up to a week. Following that, the buffer and kept at room Tissue samples may also be stored in Queen’s lysis

week. C freezer within one sferred to a -80even in a closed carboy and so tubes should be tran

C freezer. Liquid nitrogen will sublimate, nitrogen until tubes can be transferred to a -80should be stored in cryotubes in liquid cumbersome to take into the field. Frozen samples

itrogen is necessary and carboys are a field laboratory setting since access to liquid n, sample collection must be performed in (Seutin et al., 1991). If samples are to be frozen

ned from samples in lysis buffer significant amount of good quality DNA can be obtain tissue is cryogenically frozen, but a temperatures. The DNA yield is slightly higher whe

ffer or at extremely low Tissue samples must be stored either in a lysis bu

°°

°

Queen’s lysis buffer (if using) 1.5 ml microcentrifuge tubes each containing 1.0ml

Cryostore tubes (if using liquid nitrogen) Liquid nitrogen (if using)

one for Bactine 2 small dishes, one for waste isopropyl alcohol andBactine antibiotic spray Bunsen burner (or other flame source) 70% isopropyl alcohol in squirt bottle Fine-tipped forceps or tweezers

s also work well) Sharp dissection scissors (new, unused nail scissorDishtowels Fine permanent black marker

zed materials) (See Appendix 2 for suggested suppliers of speciali

Toe clipping

14

and heat sterilize between each cut. 7) Rinse scissors and forceps in isopropyl alcohol C. above 20

g within 2 minutes at temperatures for a further 30 minutes. Most turtles begin movinent is not regained, observe the turtle bleeding or swelling is observed or if normal movem

ent is regained in the limb. If swelling at the incision site and that normal movem ensure that there is no bleeding or 6) Observe the turtle at intervals for 5 minutes to

ssary. 5) Repeat procedure for the other hind foot if nece) or in storage box. storage tube and place in liquid nitrogen (if using a secure container. Tightly cap the 4) Dip the foot in Bactine. Place the hatchling inface, it is still usable. may fall. If the toe should fall on an unclean surtick to the scissors after clipping, and over a clean piece of paper as the toe will often s

d forceps (if necessary). Hold the foot immediately into the storage tube using scissors antle’s foot). Cut the toe and place it stop you from cutting the toe any closer to the tursecond knuckle (the toe webbing will down the toe until the scissor blades are past the rs, isolate a single toe and gently move forefinger. Dip the foot in Bactine. Using scisso

with toes splayed over your of the hind feet between your thumb and forefinger r palm. Gently, but firmly, grip one hind feet towards your fingers and head towards youatchling in non-dominant hand with 3) Dry hatchling thoroughly with dishtowel. Hold h

paper after sterilizing. ore scissors and forceps on clean piece of alcohol and briefly hold in flame to sterilize. St

rinse scissor blades and forceps in tube. Dispense small dish of Bactine. Thoroughly urtle ID on the cap of the tube. Open tube with fine permanent black marker. Write the t

e frosted area on the side of the storage 2) Write the turtle ID and date of collection in thcomments.

her name, recovery time of turtle and stage (e.g. hatchling), date of collection, researc Columns should include: turtle ID, life 1) Create a data sheet to track tissue collection.

Method

moved from each hind limb. painted turtles and that a maximum of one toe be reemoved only from the hind limbs of the hind limbs. Hence, we recommend that toes be rpainted turtles dig their nests using courtship and mating success. Furthermore, female these missing digits influence depredation attempts (pers obs) it is not known how

any turtles have missing digits due to forelimbs as part of the courtship ritual. While md turtles use the digits on their adjacent digits be removed. Male and female painte

mends that not more than two Ichthyologists and Herpetologists (ASIH 2004) recomn). The American Society of from the nest or caught in their first active seaso

les (i.e., those sampled directly This method should only be used for hatchling turt

°

Tail clipping

15

individual.

and heat sterilize between each 6) Rinse scissors and forceps in isopropyl alcohol ement is regained. turtle for a further 30 minutes or until normal mov

ains retracted into the shell, observe the bleeding or swelling is observed or if the tail remno longer retracted into the shell. If swelling at the incision site and that the tail is

ensure that there is no bleeding or 5) Observe the turtle at intervals of 5 minutes to ) or in storage box. storage tube and place in liquid nitrogen (if using

a secure container. Tightly cap the 4) Dip the tail in Bactine. Place the hatchling inon an unclean surface, it is still usable.

ipping, and may fall. If the tail should fall tail clip will often stick to the scissors after clhling over a clean piece of paper as the scissors and forceps (if necessary). Hold the hatc immediately into the storage tube using Bactine. Cut a ½ cm piece of the tail and place it

from the tip. Dip the tail in between your thumb and forefinger approximately 1cmm. Gently, but firmly, grip the tail feet towards your fingers and head towards your palatchling in non-dominant hand with 3) Dry hatchling thoroughly with dishtowel. Hold h

clean piece of paper after sterilizing. rilize. Store scissors and forceps on forceps in alcohol and briefly hold in flame to ste Thoroughly rinse scissor blades and tube. Open tube. Dispense small dish of Bactine. urtle ID on the cap of the tube. Open tube with fine permanent black marker. Write the t

frosted area on the side of the storage 2 Write the turtle ID and date of collection in thecomments.

her name, recovery time of turtle and stage (e.g. hatchling), date of collection, researc Columns should include: turtle ID, life 1) Create a data sheet to track tissue collection.

Method

Contact: Carol Ritland, versity of British Columbia. 1) Genetic Data Centre, Department of Forestry, Uni

Possible laboratories

ablished laboratory. We recommend that this work be contracted to an estized equipment and skills. DNA extraction and genetic analyses require special

Testing guidelines

16

[email protected]

Website: Fax (604) 822-9102 Telephone (604) 822-3908 or (604) 822-1543

http://www.forestry.ubc.ca/gdc/

s. are highly polymorphic (variable) in painted turtleon painted turtle DNA and some loci terrapins (Hauswaldt and Glenn, 2003) amplify well

d one set developed for diamondback (Osentoski et al., 2002 and Libants et al, 2004) anites developed for Blanding’s turtles especially painted turtles. Two set of microsatell

d genetic markers for turtles, through the maternal line). There are few publisheDNA (mitochondrial DNA, inherited (single nucleotide polymorphisms or repeats) and mt

peated fragments of DNA), SNPs investigated using microsatellites (small highly re among species may be Relatedness among individuals, among populations or

one day). NA may be obtained from the kit in chemicals and takes up to three days to complete (D

for tissue samples, however it uses toxic extraction is an alternative, very reliable method om tissue. Phenol-chloroform Qiagen DNeasy kit also produces high-quality DNA fr

DNA in a relatively short time. The to use and produce reliable amounts of high-qualityy kit and the GenElute kit are simple commercially available kits. Both the Qiagen DNeas

g any number of DNA may be extracted from turtle blood samples usin

communicating with the contracted laboratories: oses of effectively The following information is included for the purpNote:

work. ave a laboratory that does genetic 3) The Ministry of Agriculture and Lands may also h

am not a government worker. e information, which I cannot since I have to gain access to their website to provide mor

ey may be a good option. I would among branches of the provincial government, but th work or the nature of the relationship I am not sure whether they are set up to do genetic

nalytical Laboratory 2) Government of British Columbia Research Branch A

mtDNA and SNPs, among other genetic markers. ic testing using microsatellites, The genetic data centre accepts contracts for genet

: 82-90. 69gy samples for DNA analysis. Canadian Journal of Zoolorvation of avian blood and tissue Seutin, G., White, B.N., and Boag, P.T. 1991. Prese

149. : 147-2002. Molecular Ecology Notes Emydoidea blandingiiBlanding's turtle,

atellite loci from the C.R. 2002. Isolation and characterization of microser, M., Herman, T.B., and Hughes, Osentoski, M.F., Mockford, S.W., Wright, J.M., Snyd

: 300-302. 2004. Molecular Ecology Notes blandingiiEmydoidea loci from cross-species amplification of seven microsatellite

Congdon, J.D. 2004. Isolation and Libants, S., Kamarainen, A.M., Scribner, K.T., and

: 174-176. ). Molecular Ecology Notes Malaclemys terrapinterrapin (e DNA loci from the Diamondback Hauswaldt, J.S. and Glenn, T.C. 2003. Microsatellit

: 457-473. Biology and Evolution he Testudines. Molecular variability and reduced microevolutionary rate in t

dence for low genetic Mitochondrial DNA evolution at a turtle’s pace: evi. d Bermingham, E. 1992Avise, J.C., Bowen, B.W., Lamb, T., Meylan, A.B. an

http://www.asih.org/files/hacc-final.pdf mudio. Members: Jacobson, E.R., Lillywhite, H.B. and K. Za

Chair: Beaupre, S.J., Herpetological Animal Care and Use Committee (HACC) ed, Rev by ndearch, 2amphibians and reptiles in field and laboratory res

ts. 2004. Guidelines for use of live American Society of Ichthyologists and Herpetologis

References

17

9

3

Blood lysis cards Blood lysis buffer

s Appendix 1: Cost comparison of blood storage method

18

Item required Cost/sample Item required Cost/sample 1.5 ml microcentrifuge tube $0.07 FTA microcard $3.02 lysis buffer pennies Multi barrier pouch (sm) $0.84 storage boxes $0.25 desiccant $0.07

Laboratory costs Extraction kit GenElute or Dneasy $2.48 0.5ml PCR tubes $0.09 Purification reagent $0.11 TE buffer pennies

$2.80Total $4.13

for DNA extractionrequire a specialized hole punch

: Blood lysis cards also Note

Supplier Catalog numberItem

Appendix 2: Suggested suppliers

Supplier number Cost Blood sampling

4 $39.90/100 bevel length ½”, size 27 gauge Sigma-Aldrich Z19238B-D Precisionglide syringe needles,

ma- Aldrich Z192090 $50.80/100 Polypropylene tuberculin syringe 1.0ml capacity Sig Cell lysis buffer method

ntific 02-681-284 L-510-GRD-FIS 33.27/500 Fisherbrand 1.5ml microcentrifuge tubes Fisher Scie B4183-1CS 152.50/25 boxes Nunc 25-cell storage box for Cryoline Sigma-Aldrich

ich NA2010-1KT 173.70/70 preparations GenElute blood genomic DNA kit (70 prep) Sigma-AldrDNeasy blood and tissue kit (50 prep) Qiagen 69504 168.00/50 preparations

Cell lysis card method Z719773-100EA 302.00/100 cards Whatman FTA technology FTA microcard Sigma-Aldrich

715-111 WB100036 83.70/100 Whatman barrier pouch (small) Fisher Scientific 05-dDrierite dessicant 3g bag WA Hammond Drierite Co Lt 60013 52.47/750

1EA 274.50/500ml Whatman purification reagent Sigma-Aldrich Z719978- Tissue in liquid nitrogen

494.50/500 1.0 ml Nunc cryotube vials Sigma-Aldrich V7384-1CS

the analyzing laboratory. laboratory charges. This should be negotiated with

purification reagent may be included in any : Cost of DNA extraction kits or the Whatman Note

western painted turtle monitoring at alaksen national wildlife … · 2011. 4. 28. · western...

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