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|         |                                                   |        |
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| !       | **United States Department of the Interior**      | ![     |
| [](medi |                                                   | ](medi |
| a/image | **FISH AND WILDLIFE SERVICE**                     | a/imag |
| 1.png){ |                                                   | e2.png |
| width=" | Division of Ecological Services                   | ){widt |
| 0.98888 |                                                   | h="0.8 |
| 8888888 | 9014 East 21^st^ Street                           | 375in" |
| 8889in" |                                                   | hei    |
| he      | Tulsa, Oklahoma 74129-1428                        | ght="1 |
| ight="0 |                                                   | .0in"} |
| .988888 | 918/581-7458 / (FAX) 918/581-7467                 |        |
| 8888888 |                                                   |        |
| 889in"} |                                                   |        |
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|         |                                                   |        |
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**American Burying Beetle *Nicrophorus Americanus***

**Survey Guidance For Oklahoma**

**Updated June 13, 2007**

*[Introduction]{.underline}*

This guidance is provided to aid in designing and conducting surveys for
compliance with the Endangered Species Act. Surveys for ABBs that are
primarily conducted for research also should be coordinated with the
Service. Baited pitfall traps are the most effective method known for
surveying for ABBs (Creighton *et al.,* 1993; Service, 1991; Bedick *et
al,.* 2004). These methods as outlined below have proven to be
successful in capturing the ABB. Following this guidance should help to
ensure the validity of survey results. Further, data gathered using this
guidance will allow for comparison of results between surveys.

*[Time for Surveys]{.underline}*

The Service determined that trapping between May 20 and September 20
would adequately document presence/absence of ABBs. Surveys should begin
no earlier than May 20 and end no later than September 20, but these
dates may vary among states and will need to be verified for each state.
Each night during a survey effort where the night-time ambient
temperature falls below 60ºF an additional night of surveying should be
conducted.

ABBs are active from May 20 through September 20 in Arkansas and
Oklahoma. This is based on the past ten years of weather data in eastern
Oklahoma and western Arkansas. May 20 is typically the day commencing
the onset of a five day period were nighttime ambient temperatures
remain 60 degrees Fahrenheit \[ºF\] or higher, and September 20 is the
first day of a five day period with temperatures below 60ºF. However,
there is the potential for a night or two where the temperature drops
below 60ºF but not for five nights or more.

*[Timing]{.underline}*

The Service has not restricted specific time frames when ABB survey must
be conducted between May 20 and September 20 timeframe.

Capture rates for ABBs are highest from mid-June to mid-July and again
in mid-August. Both Bedick *et al.* (2004) and the U.S. Fish and
Wildlife Service (Service) (1991) reported an increase in capture rates
during a certain time of the ABBs active season and during the night.
Both indicated that the period from mid-June to mid-July was a peak ABB
capture timeframe. Bedick *et al.* (2004) reported that a peak in
capture success also occurred in mid-August. Service (1991) reported
that during late July ABBs were easy to attract to carrion bait but were
difficult to capture in pitfall traps.

Surveying during the above peak timeframes would result in the greatest
potential for trapping success and the Service would prefer that surveys
be conducted during these periods. However, we understand that limiting
surveys to these time periods is too restrictive and therefore
conducting surveys during these time periods is not a requirement.
Further, baited pitfall trapping is only a method to determine
presence/absence and not an abundance or population survey.

*[Timeframe a Survey is Valid]{.underline}*

Based on the annual cyclic nature of ABB populations, survey results
should only be valid for one year from the last date of the survey.
After one year, data indicates that ABBs likely have moved and numbers
have fluctuated within a given area.

Much of the long-term information concerning the life history of the ABB
pertinent to Oklahoma has come from work done at Fort Chaffee, Arkansas,
Weyerhaeuser Company lands in Oklahoma, and Camp Gruber in Oklahoma.
While the land use at Fort Chaffee, AR; Weyerhaeuser, OK; and Camp
Gruber, OK is different among each entity, each entity maintains a
relatively consistent land use pattern of its own through time. However,
Hiott and Schnell (2003) reported the number of ABBs captured and the
location of high density ABB concentrations varies annually at each
site. This trend indicates ABBs are annually cyclic, where there are
high numbers and abundance one year, followed by a decline in numbers
the following year, and repetition of the cycle over time. In addition,
each year they reported that the high concentration appeared to shift
annually throughout the sites. Further, the ABB is an annual species
(living for only one year) and the following year's numbers are
dependent upon the reproductive success of the previous year.

False negatives are possible outcomes of with ABB surveys. The Service
(1991) reported that during late July ABBs were easy to attract to
carrion bait but were difficult to capture in pitfall traps. Standard
transects on Camp Gruber that resulted in ABB captures in one year
showed no capture of ABBs in another year. Surveys conducted in a given
area have resulted in ABB captures during one survey but surveys
conducted in the same given area and the same active season have
resulted in negative ABB captures.

*[Survey effort radius]{.underline}*

Considering ABBs mobility, small size, recorded movement distances, and
distance from which they can detect carrion, a presence/absence survey
was effective only over 5 miles and that this erred on the side of the
species.

An ABB may move as little as 0.15 mile per night to a maximum of 6.2
miles in 6 nights (1.03 miles per night), with a mean movement distance
of 0.52 mile per night (Schnell and Hiott 1995, ­2003). Bedick *et al.*
(2004) reported average nightly movements of 0.62 mile with the large
proportion (85%) of recaptures moving distances of 0.31 mile per night.
Creighton *et al.* (1992) reported that individual ABBs moved over 4
miles in only a few days. Creighton and Schnell (1998) reported an
average nightly movement of 0.76 mile, and maximum distances of four
miles in five nights (0.8 mile per night) and 6.2 miles in 6 nights
(1.03 miles per night), a minimum distance of 0.15 mile in one night,
and a mean nightly movement of 1.67 miles.

While this data could be interpreted to imply that an ABB could move 95
miles \[0.62 (mean nightly movement) X 154 days (May 20 -- September
20)\] during the active season, the Service does not believe this is an
accurate interpretation. Mark and recapture data at Camp Gruber and Fort
Chaffee did not find any ABBs that moved between these installations, a
distance of about 54 miles (Schnell and Hiott, 1997-2003). Even if ABBs
moved such long distances, the Service assumes it is unlikely ABBs move
in such a consistently linear direction.

The group of ABBs at Camp Gruber has persisted for at least 11 years.
While the numbers and high density areas of ABBs have changed annually,
indicating ABBs are a cyclic species (Schnell and Hiott, 2003), they
appear to be self-sustaining. The Service assumed it was not likely that
all, the majority, or even a single ABB on these installations moved
from 54 to 95 miles away from Camp Gruber. Further, for a species to
survive it cannot exert more energy than it consumes and would only
expend the minimal amount of energy needed to secure necessary
resources.

Considering the available data regarding ABB movements and life history
patterns, and the need for establishing an effective trapping area, the
Service believes the 5 mile radius standard accurately represents the
best available scientific information.

*[Minimum Survey Effort]{.underline}*

To effectively determine presence or absence of ABBs in a given area
surveys need to be conducted for a minimum of 3 consecutive nights \[or
24 trapnights, 8 traps (or 1 transect) x 3 nights = 24\].

Transects should be spaced at 0.5 mile increments across the project
area until the entire project area is within the effective trapping area
of at least one transect and each one of these transects should be
deployed for the same 3 consecutive nights.

Any night during the survey when the ambient temperature drops below
60ºF, the results should not be included in survey results (unless ABBs
are captured although this is unlikely) and an additional night of
surveying is required.

Any night during the survey when there is more than rain 1/2 inch of
rainfall, the results should not be included in survey results (unless
ABBs are captured although this is unlikely) and another night of
surveying is required. For previous nights weather data the Service
recommends consulting: <http://www.nws.noaa.gov/climate/>

ABB surveys are not to be conducted in known high density areas when
rain is forecasted to occur within the project area during the
anticipated survey. Surveys may be conducted in low density areas if
there is less than a 25 percent chance of rain within a 36 hour period
prior to sampling. High density areas are defined as: average 1 or more
ABBs/trap night/transect; transects consist of 8 traps spaced 66 feet
apart and surveyed for three consecutive nights; area includes a 0.5 mi
polygon extending around the transects with an average of 1 or more
ABBs/trap night/transect. Traps should be closed during heavy rains,
particularly if rain does not begin until well into the night after ABBs
have become active and may already be in the traps. Rain that is medium
to heavy, or even light rain that continues all night, could reduce the
likelihood of capturing ABBs. Defining heavy rain can be somewhat
subjective, so the permittee will have to use professional judgment to
some degree.

Creighton *et al.* (1993) and the Service (1991) recommend surveying for
a minimum of 3 consecutive nights to adequately determine the presence
or absence of ABBs in an area. Bedick *et al.* (2004) recommends a range
of 3 to 5 nights, preferably 5 nights. However, after 3 consecutive
nights of trapping, surveyor observations report trapping success
declines. If ABBs have not been captured within the 3 consecutive nights
of surveying then ABBs are unlikely, although not impossible, to be
captured (Personal communication with Ana Hiott, 2003) with additional
trapnights.

Specific data is lacking regarding the minimum number of trapnights or
transects necessary to determine presence or absence of ABBs during 3
nights of trapping. However, based on the movement data described above,
the effective transect area is a 0.5 mile radius. So to effectively
survey a particular area, transects should be spaced at 0.5 mile
increments across the project area until the entire project area is
within the effective trapping area of at least one transect and each one
of these transects should be deployed for the same 3 consecutive nights.
Keep in mind that a transect does not have to be linear.

For projects with less than a 0.5 mile radius from the center to any
point along the perimeter, a minimum of 1 complete transect (8 traps
spaced 66 feet apart) is required. One transect with 8 traps for 3
nights results in 24 trapnights (the number of traps times the number of
survey nights). Again, the placement of the transect does not need to be
linear

*[Minimum project acreage]{.underline}*

According to our analysis of ABB density estimates, projects which
disturb less than 1.2 acres would have, on average, no more than a one
percent chance of impacting an individual ABB. In addition, the Service
evaluates numerous other factors including the project: type,
construction/installation duration, permanency, location, land use,
implementation methods, habitat, and time/season.

Using recently collected survey data, the Service's Oklahoma Ecological
Services Field Office derived densities of ABBs in their known range
within Oklahoma. We used all known survey records to determine the total
number of ABBs captured and total transects deployed per survey. In
using this information, we assumed all captures were obtained using the
methodology described by Creighton, *et al*. (1993) unless stated
otherwise. Data that were not collected using the Creighton *et. al*.
(1993) methodology were excluded from the analysis.

The Service then estimated the area each transect would effectively
trap. Creighton, *et al*. (1993) determined, based on known movements of
ABBs, that transects do not need to be spaced any closer than 0.5 miles.
Past and ongoing research demonstrates this trapping recommendation is
still appropriate. Schnell and Hiott (1997-2003) annually determined the
average nightly movements of the ABB, using marked individuals over a
nine-year period at Camp Gruber to be 0.62 miles. The smallest average
nightly movement for any given active season over that same period was
0.52 miles. Consequently, we believe each transect effectively traps
beetles from an area of 0.5 mile. As such, the effective trapping area
(ETA) for one transect is about 153.5 acres. Using the ETA and number of
ABBs collected, we estimated average ABB densities to be 0.0084
ABBs/acre for their known range in Oklahoma. The Service then used a
standard z test to determine the probability of encountering an
individual ABB in a given area. We determined that disturbance of less
than 1.2 acres would have, on average, no more than a one percent chance
of impacting an individual ABB.

*[Transect Placement]{.underline}*

Transects should be placed in a configuration that best represents the
different habitat types present in the survey area and on the highest
spots in the survey area.

American burying beetles are feeding habitat generalists (Creighton *et
al,* 1993). Consequently, it is recommended that transects be placed in
all the different habitat types present in a survey area. Bedick *et
al.* (2004) reported ABBs were significantly more attracted to traps
placed on ridges than those placed in valleys. They believed that
increased ABB captures on ridge tops may be a result of increased odor
movement, thus increasing the possibility of detection by ABBs.

*[Ants]{.underline}*

Traps should not be placed within 23 feet (7 meters) of ant hills. If
ants are discovered in a trap it should be relocated at least 23 feet
away. Ants can swarm and kill an ABB that is trapped in a pitfall trap
(Creighton *et al,.* 1993 and Bedick *et al,.* 2004).

*[Transect spacing]{.underline}*

Transects should be spaced a minimum of 0.5 mile apart on small to
medium sized projects and a maximum of 1 mile apart on large projects to
achieve reliable survey results.

Bedick *et al.* (2004) recommended that 0.31 mile be used as a minimum
transect spacing for traps with large bait. Creighton *et al.* (1992)
concluded that transects less than 0.5 mile apart were not required. In
large sample areas, Creighton *et al.* (1992) stipulated that transects
can be located as much as 1 mile apart. The Service's rationale
considered these recommendations and the ABB movement data discussed
previously.

*[Transect]{.underline}*

A transect is defined as 8 pitfall traps, spaced 66 feet (20 meters)
apart (Figure 1), for a total length of 460 feet (140 meters). This
definition is based on the transect protocol described in Creighton *et
al.* (1993) and Bedick *et al.* (2004) which all utilize or recommend
the same type of transect. Transects are not limited to linear
arrangements, they can be aligned to suit the shape of the project area
for which you are surveying. For example, if a project is kidney bean
shaped, transects can be curved to fit a kidney bean shape. However, the
above described quantity of traps per transect and the spatial
arrangement (traps 66 feet apart) of traps must be followed.

*[Traps]{.underline}*

The Service recommends using baited pitfall traps for ABB
presence/absence surveys. Each trap consists of a trap cup, bait cup,
wire, and cover. The traps are designed to attract ABBs with the carrion
bait, keep captured ABBs alive, and prevent escape until traps are
checked the following morning. The ABB flies toward the bait odor, lands
near the trap, and crawls under the cover and into the trap cup. Once in
the trap, the slick sides of the cup prevent the ABB from crawling out,
and the cover and suspended bait cup keep the ABB from flying out of the
trap cup. Figures 2 and 3 display the baited pitfall trap setup. If the
utilization of other trap design and equipment is proposed this must be
coordinated with and approved by the Service.

Traps in the form of 24 oz (0.7 L) plastic cups (similar to Solo cups)
have proven effective. The Service recommends that the trap cup consist
of two 24-oz Solo cups. Cups must have smooth sides, free of any texture
or ridges to prevent ABBs from escaping. American burying beetles can
climb a surface with textured/ridged sides but cannot climb a smooth,
slick surface. The 2 cups are stacked together and placed in an
appropriate sized hole in the ground. Stacking the cups one inside the
other facilitates removal of trapped insects. The top cup can easily be
pulled out of the ground and replaced while the second cup remains in
the ground to maintain the integrity of the hole. The lip of the cups
should be 0.5 to 0.75 inch above ground level. This prevents water
runoff from filling the cup. A 1 to 2 inch squared piece of wetted
sponge soil should be placed in the bottom of the trap cups. This has
been shown to decrease mortality of ABBs from desiccation.

The bait cup consists of a plastic cup about 5 -- 6 ounces (20
milliliters) in size. Examples of bait cup types that have proven
effective are cups similar to those used in restaurants for carry out of
salad dressing or styro-foam coffee cups with the top portion cut off
leaving only the bottom 1 inch of the cup. The type of bait cup used is
not as important as the trap cup, however the size of the bait cup is
important. The bait cup should be large enough to hold the proper amount
of bait but small enough to be suspended over the trap cups and still
allow ABBs access to the trap cup. Bait suspended over the pitfall trap
via skewer or similar device is not recommended. The bait dries out and
the odor emitted is strongly weakened.

Wire is used to secure and suspend the bait cup over the trap cup. To
accomplish this, the wire needs to be hand malleable but sturdy enough
to support the bait cup. The wire is inserted into one side of the bait
cup near the top and pushed all the way through the cup and out the
other side of the cup; about 3 inches of wire should extend on either
side of the exterior of the bait cup. Bend wire down and push into the
ground to suspend and secure the bait cup over the trap cup. The reason
for separate bait and trap cups is to protect the ABB from coming in
contact with the bait. As the bait liquefies or becomes gummy ABBs could
be harmed if they come into contact with the bait.

The cover should be hard, not transparent or opaque, weighted or secured
to the ground, roughly 6 inches in depth, about 12 inches in diameter at
its widest point, and raised off the ground about 1 to 2 inches. A hard,
plastic dome structure has proven effective at protecting the trap (e.g.
inverting nursery plant containers with holes on the side, or small gray
paint buckets). A hard structure, secured to the ground is needed to
prevent scavenger and water access into the trap cup, and to provide
shade to captured insects to avoid desiccation. Scavengers absconding
with bait is common and potentially results in harm to ABBs. In
addition, bait loss during trapping reduces the effectiveness of the
trapping effort and therefore ABB capture success. The structure should
be raised of the ground about 1 inch or holes should be cut along the
lip of the container to allow ABBs to crawl into the trap. At least 3
holes need to be provided for ABB access. In lieu of a plastic dome
structure, other similar structures can be used. For example, a piece of
wood raised on legs can be used. Shingles or other such flimsy materials
are not to be used.

In areas where scavengers are a significant problem wire mesh can be
installed between the pitfall trap and the cover. The piece of wire mesh
should be a minimum of 6 inches squared and mesh size should be at least
1 inch. The wire mesh should be secured to the ground with stakes, and a
hard cover will still need to be used.

Exposure to full sunlight and temperatures over 77ºF for even a few
hours can result in ABB mortality (Service, 1991). According to Bedick
*et al.* (2004) wetted soil in the bottom of the trap helped maintain
high humidity. This improved the longevity of ABBs and therefore
decreased the mortality of ABBs from desiccation. They found this was
feasible to use with stacked cup traps. Other permittees have used a 1
to 2 inch squared piece of wetted sponge in the bottom of traps.

Bedick *et al.* (2004) found the primary cause of ABB mortality was
drowning due to traps flooding with water. However, on multiple
occasions, floating and apparently dead beetles were removed from traps,
and they subsequently recovered after 10 to 20 minutes. Consequently,
ABBs that appear dead or are lethargic should be held for at least 20
minutes to determine actual condition. Monitored ABBs should be placed
in a roomy, open container, with air circulation, out of direct
sunlight.

Bedick *et al.* (2004) evaluated six types of pitfall traps for
capturing ABBs ranging from stacked cups, to PVC pipe, to buckets. They
determined buckets with a dome cover had the greatest trapping success
because it allowed for the largest piece of bait. However, they realized
that this was not the most practical or economically feasible method to
implement and rated stacked cups as the second best trapping method.
Creighton *et al.* (1993) also determined that stacked cups as baited
pitfall traps proved effective at capturing ABBs and recommended this
method.

Creighton *et al*. (1993) recommended using traps that are smooth, free
from any bump or ridge to prevent ABBs from climbing out of the trap.
Metal cans should be avoided because as soon as any rust appears, ABBs
may be able climb out of the trap (Service, 1991). Creighton *et al.*
(2004) also recommended the lip of the cup be flush with the ground.
However, Bedick et al . (2004) and Hiott (2002 personal communication)
recommend the lip of the pitfall trap be a 0.5 to 1 inch above the
ground surface to prevent water runoff from filling up the trap. Bedick
*et al.* (2004) took additional measures by building a berm around the
raised portion of the cup.

Creighton *et al*. (1993) and Bedick *et al.* (2004) recommend
suspending bait cups over trap cups to lure ABBs, and prevent contact
between ABBs and the bait. While carrion beetles are seemingly
well-adapted to moving around in carrion, Bedick *et al.* (2004) found
that under some conditions spiracle blocking could occur. They did not
think this was a common occurrence but could potentially occur. For
example, bait placed in a small trap with a water-tight bottom can decay
into a near liquid state. Bait in this liquid form could adhere to ABBs,
harden, and then clog spiracles.

Bedick *et al.* (2004) recommends using weighted, hard plastic
rain-shields larger than the opening of the trap to protect the trap
from rain. The cover over the pitfall trap should be secured to the
ground to effectively protect the trap from rainfall and predators
(Creighton *et al.,* 1993). Bedick *et al*. (2004) also recommended
using wire mesh secured to the ground between the trap and the cover
(Figure 4). Mesh was secured to the ground with three to four bent wire
stakes, 4 inches (10 cm) in long, driven into the ground. Covers were
raised between 1-2 inches above the ground to allow ABBs access to trap.

*[Bait]{.underline}*

Any type of carrion is suitable for use as bait for ABB pitfall traps.
However, un-skinned chicken is preferred by multiple surveyors because
it is inexpensive and remains moist longer than other baits due to the
fat content of the skin. Others have successfully used liver, gizzard,
or road kill. The only requirements are that the bait must be the
appropriate size in correlation with trap size and must produce a
pungent odor that ABBs are able to detect. The appropriate size of bait
for stacked cup traps is between 0.5 to 0.7 ounces (15-20 grams). The
bait must be rotten and emit a pungent odor to be effective. Adding a
small amount of water to the bait cup is effective at prolonging the
moisture content and odor. During trapping efforts any bait that has
dried out or no longer emits a pungent odor must be replaced with new
bait.

Bedick *et al.* (2004) reported no significant difference between
capture rates of ABBs using various types of bait (Appendix 1). However,
they did find a significant difference between the sizes of bait used
and the number of ABBs captured. A large piece of bait positively
correlated with an increase in the number of ABBs captured. Bedick *et
al.* (2004) recommended that bait of 7 ounces (200 grams) be used. This
amount of bait is feasible if a bucket trap is used but not if a cup is
used. Creighton *et al.* (1993) and Service (1991) recommend that
0.5-0.7 ounces (15-20 grams) of bait be used.

Fresh bait is not an attractant to ABBs (Creighton *et al.,* 1993). To
prepare bait for use, cut into cubes about 0.5-0.7 ounces (15-20 grams)
in size. Place the cubes into a sealed container or bag. Do not fill the
container or bag completely full, as the bait rots gas pressure is
increased and the extra room is need for this expansion. Place the
container or bag in the sun for a minimum of one day (Creighton *et
al.,* 1993). If the day is relatively cool (less than 85ºF \[29ºC\]),
the bait should stay in the sun longer.

Once bait is prepared, the packaged (container or bag) bait should then
be further enclosed in a larger sealed container, such as an ice chest
or bucket with lid. The bait should be used within the next few days.
Use of the larger sealed container prevents odor from escaping into
unwanted areas and is ideal for transporting bait. The Service
recommends that the bait or any containers holding bait not be placed
inside a vehicle. You will never get the smell out of your vehicle!
Instead, place the containers in the bed of a pick-up or secure to
outside of vehicle. In addition, discarded or old bait should not be
left at or near the current trapping area. This could lure ABBs away
from the baited pitfall traps.

*[Setting and Checking Traps]{.underline}*

Each trap must be checked by 10:00 A.M. each morning. Checking traps
entails: collecting all trapped ABBs, recording and releasing other
*Nicrophorus* species; replacing any missing or dry bait, re­moistening
soil or sponge in trap bottom if needed, and replacing/resituating any
disturbed parts of the trap.

Any injured or lethargic ABBs should be released immediately. ABBs that
appear to be dead should be collected and monitored for at least 20
minutes, as described previously, to accurately determine their
condition. Any dead ABBs should be handled as described below.

The Service recommends using a 5-gallon bucket or similar container to
carry the needed equipment used during trap checking. The items you will
need to carry include: a container to hold ABBs, prepared bait, water, a
trowel for digging, tongs and the data sheet. Use a hard, plastic
container with a lid and air holes to hold ABBs collected along a
transect. Carry a small container of prepared bait (and tongs to handle
bait) to replace missing or dried bait. Carry a 20 oz plastic bottle of
water to re-­moisten the soil or sponge in the bottom of the cup trap,
and the bait. Replace or re-situate any disturbed traps (this is where
you may need the trowel to re-dig the hole for the trap). Record the
species and number of any other *Nicrophorus* captured and then
immediately release. The other *Nicrophorus* species should be released
several feet away from a transect to avoid being crushed by foot
traffic. Once all the traps along a transect have been checked, proceed
in processing all the captured ABBs either at the end of the transect or
at the vehicle.

All traps must be in place and baited by 5:00 P.M. each night. Traps
should be cleared of ABBs by 10:00 A.M. every day. Traps can be baited
at the same time they are checked each morning provided the bait does
not dry out. Since ABBs are nocturnal, there is not a risk of baited
traps capturing ABBs during the day. Exposure to full sunlight and
temperatures over 77ºF (25ºC) for even a few hours can result in ABB
mortality (Service 1991).

*[Disturbed bait or traps]{.underline}*

Predators or scavengers can cause bait loss and/or trap disturbance
during ABB surveys. This can affect the results of the trapping effort.
Disturbed traps or traps missing bait, reduces or eliminates the
effectiveness of attracting and/or containing ABBs and therefore ABB
capture success. Any negative results from these traps are not reliable.

If during a 3 night survey period, no *Nicrophorus* species have been
captured and more than 2 traps along a given transect have been
disturbed or bait taken during each night, an additional night of
trapping is to be implemented for each transect meeting this criteria.
Transects in which *Nicrophorus* have been captured, regardless of
whether or not any traps have been disturbed or are missing bait, do not
require additional nights of trapping since *Nicrophorus* species have
been confirmed detecting and finding baited pitfall traps along a
transect.

*[Processing ABBs]{.underline}*

Processing includes sexing, aging, taking measurements, marking (if
authorized) and data recording.

An efficient method to process ABBs is to collect all the ABBs from all
8 traps along a transect and then process all the ABBs at one time.
Processing of ABBs can then be completed at the end of the transect or
at the vehicle. Multiple permittees find processing ABBs at their
vehicle facilitates the task. This eliminates the need to carry all of
the equipment during trap checking, quicker processing of ABBs,
availability of a cooler if needed, and other advantages.

Clipping of the elytra is only applicable for mark and recapture surveys
and is not appropriate without specific authorization from the Service.
The survey protocol described here is to determine the presence or
absence of ABBs only. Clipping of elytra causes hemoglobin to be
secreted by ABBs. Although, there is currently no conclusive evidence
that this is a direct or indirect adverse impact to the ABB it is not
recommended. It is the Service's responsibility to err on the side of
the species. Further based on past survey reports, recaptures are highly
unlikely during such a short survey effort. If a research project is be
conducted and identification of individuals is needed then bee tags can
be used. These can be attached to the elytra with superglue. Tagged ABBs
must be monitored until glue dries (about 5 minutes) to ensure wings are
not glued together and they are able to fly.

Measuring the pronotum is voluntary, however the Service would
appreciate the collection of this data. Measuring of the pronotum should
be done with calipers, preferably digital.

*[Identification and Other Nicrophorus Species]{.underline}*

There are 6 other *Nicrophorus* species in Arkansas and Oklahoma that
resemble the ABB. The ABB can usually be distinguished from other
*Nicrophorus* species by the large orange-red spot on its pronotum (body
segment between the head and abdomen) and on the frons (Figure 5). No
other mature *Nicrophorus* species has an orange-red marking covering
the pronotum and frons (Figure 5). However, newly eclosed (within 2
weeks after emerging from the ground) *N. orbicollis* can have a burnt
orange marking on the pronotum. Another characteristic to look for on
the ABB to distinguish it from *N. orbicollis* is red-orange frons. The
*N. orbicollis* has black frons. A description of the other
*Nicrophorus* species in Oklahoma and an identification key is located
in Appendices 2 and 3, respectively. In addition, photos of other
*Nicrophorus* species in Oklahoma are available on our website for
comparison.

*[Sexing]{.underline}*

The sex of ABBs can be distinguished based on the orange-red marking
located between the frons and mandibles on the head. These markings are
rectangular on males and triangular on females (Figure 6).

*[Aging]{.underline}*

Adults that have pupated during the current active season are known as
newly eclosed. They can be distinguished from the previous years young
by their softer bodies and more shiny appearance (Creighton *et al.,*
1994). The orange-red pronotum appears to be lighter and more orange in
color in newly eclosed adults. Older adults often are missing body
parts, especially legs or antennae. In addition, the mandibles of older
adults appear to be a bit more worn at the tip.

*[ABB Release]{.underline}*

ABBs should be released along transects where they were captured or
within about 500 feet of the transect. Further, ABBs should be released
a minimum of about 100 feet away from vehicle or foot traffic and
outside of the pathway of vehicle and foot traffic. Captured ABBs should
only be held for a maximum of 30 minutes, preferably much less than
this. ABBs held for longer than 10 minutes should be placed in a hard,
plastic container with a damp sponge and then the container placed in an
iced cooler. American burying beetles are sensitive to prolonged heat
exposure.

*[Reporting]{.underline}*

The Service has prepared a standard "ABB Survey Data Form"(Appendix 4).
Use of this form ensures that all of the needed data is recorded by all
permittees. This form is to be completed for each transect, each night
during a survey effort. Copies of all forms are to be submitted to your
local Ecological Services field office.

In addition, an "ABB Summary Report" (Appendix 5) is to be completed for
each survey effort (Appendix 4). This is to be submitted in electronic
and hardcopy format to your local Ecological Services Field Office along
with the corresponding "ABB Survey Data Forms". A description of the
required fields to complete in the "ABB Summary Report" is provided in
Appendix

*[Accidental Death of ABBs]{.underline}*

The handling of all endangered species is strictly regulated by the
Service. All accidental mortalities of ABBs must be accounted for and an
'*ABB Accidental Death Form'* must be completed for each individual
specimen and submitted to the Service within 14 calendar days to the
appropriate Ecological Services Field Office. An '*ABB Accidental Death
Summary Report'* must be completed by October 15 of each year in
electronic and hardcopy formats and submitted to your local Ecological
Services Field Office and Regional Permit Coordinator.

Dead ABBs should be placed in cotton within a sealable, rigid container
to prevent jostling of the ABB causing limb and antennae damage. Each
specimen must have a unique alphanumeric name assigned. This
alphanumeric name should be the first letter of the first 2 words of the
permittee company or individual (e.g. Acme Company, first dead ABB =
AC001). A label with the date found dead, permittee, legal description
(down to quarter section at least), and specimen alphanumeric name
should be placed inside each container to ensure future identification.
Only place one ABB specimen per container to avoid mixing up specimens.
Place the container on ice until the ABB can be prepared. Dead ABBs are
to be submitted to the Service or a Service approved facility.

*[Protocols and Forms]{.underline}*

All forms can be downloaded from the Oklahoma Ecological Services Field
Office's website
\<http://www.fws.gov/southwest/es/oklahoma/beetle1.htm\>.

**References**

Bedick, Jon, Brett Ratcliffe, and Leon Higley. 2004. A new sampling
protocol for the

> endangered American burying beetle, *Nicrophorus americanus* Olivier
> (Coleoptera:
>
> Silphidae). The Coleopterists Bulletin 58(1):57-70.

Creighton, Curtis, Mark Lomolino, and Gary Schnell. 1993. Survey methods
for the American burying beetle (*Nicrophorus americanus*) in Oklahoma
and Arkansas. Unpublished.

Creighton, Curtis, and Gary Schnell. 1998. Short-term movement patterns
of the endangered American burying beetle *Nicrophorus americanus*.
Biological Conservation 86:281-287.

Schnell, Gary, Ana Hiott. 1995. 1995 Annual report of trapping and
relocation activities

concerning the endangered American burying beetle (*Nicrophorus
americanus*). Sam

Noble Oklahoma Museum of Natural History, University of Oklahoma.
Unpublished.

Schnell, Gary, Ana Hiott. 2003. 2003 Annual report of trapping and
relocation activities

concerning the endangered American burying beetle (*Nicrophorus
americanus*). Sam

Noble Oklahoma Museum of Natural History, University of Oklahoma.
Unpublished.

U.S. Fish and Wildlife Service. 1991. American burying beetle
(*Nicrophorus americanus*) recovery plan. Newton Corner, Massachusetts.
80 pp.

This guidance was developed from the above references, U.S. Fish and
Wildlife Service's July 14, 2005, "ABB Survey Guidance" and a U.S. Fish
and Wildlife Service Working Group on May 6, 2004. The Oklahoma
Ecological Services Field Office, in coordination with other Field
Offices, update this survey protocol as necessary due to new findings.
The purpose of this guidance is to streamline and update American
burying beetle survey recommendations among the Arkansas, Oklahoma,
Kansas, and Arlington, Texas Field Offices.

**AMERICAN BURYING BEETLE SURVEY FORM\
**

Time:\_\_\_\_\_ Date:\_\_\_\_\_\_\_ Transect #:\_\_\_\_\_\_ Survey
Night: 1 2 3 4 5 6 State:\_\_\_\_\_

> D/ M / Y

Survey Company: \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Surveyor:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Landowner:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_

County:\_\_\_\_\_\_\_\_\_\_\_ Legal
Description:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ General
Location:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_

> (Sec Township Range) (state mngt area, refuge, etc)

Decimal
Degrees:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_/\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Coordinate System:\_\_[NAD83]{.underline}\_

Vegetation Type:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Primary Soil Type:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
\_\_\_\_\_\_\_

> (Prairie, woodland, forest, pasture) (Refer to County Soil Survey)

Temp: Min\_\_\_\_\_\_ Max\_\_\_\_\_\_ °F Wind:\_\_\_\_\_\_\_mph Cloud
Cover:\_\_\_\_\_\_\_\_\_\_\_\_\_%

Population: Wild\_\_\_ Reintroduced\_\_\_ Relocated\_\_\_ (If
Relocated/Reintroduction See Appropriate Data Form)

  ----------- -------------- -------------- -------------- -------------- -------------- ------------- -------- ----------
  Trap No.    *americanus*   *orbicollis*   *tomentosus*   *pustulatus*   *marginatus*   *carolinus*   *sayi*   *Other
                                                                                                                insects*

  1\. P GTI                                                                                                     
  GDU                                                                                                           

  2\. P GTI                                                                                                     
  GDU                                                                                                           

  3\. P GTI                                                                                                     
  GDU                                                                                                           

  4\. P GTI                                                                                                     
  GDU                                                                                                           

  5\. P GTI                                                                                                     
  GDU                                                                                                           

  6\. P GTI                                                                                                     
  GDU                                                                                                           

  7\. P GTI                                                                                                     
  GDU                                                                                                           

  8\. P GTI                                                                                                     
  GDU                                                                                                           

  Totals                                                                                                        
  ----------- -------------- -------------- -------------- -------------- -------------- ------------- -------- ----------

List each individual American burying beetle captured below and complete
the appropriate columns.

  --------- ---------- ------------ --------- --------- ----------- --------------- ---------- ------- ----------- ------------
  **ABB**   **Male**   **Female**   **Old**   **New**   **Age       **Recapture**   **Newly    **Tag   **Death**   **Pronotum
                                                        Unknown**                   Marked**   \#**                Width**

  1                                                                                                                

  2                                                                                                                

  3                                                                                                                

  4                                                                                                                

  5                                                                                                                

  6                                                                                                                

  7                                                                                                                

  8                                                                                                                

  9                                                                                                                
  --------- ---------- ------------ --------- --------- ----------- --------------- ---------- ------- ----------- ------------

Total Number of Years
Surveyed:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_

Comments:\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_

Max/Min temp for previous 24-hour period:

Other weather data refer to current conditions.

Date and time refer to when trap checked;

P=Bait present; GTI=Bait gone trap intact; GDU=Bait gone trap dug up;

OLD=breeding adult; NEW=newly eclosed adult; UNK=age cannot be
determined.

Newly marked males and females refers to color, number of bee tag, and
age of beetle (e.g. R54\[old\]).

Recaptures refer to color and number of bee tag on beetles that have
been previously marked.