Experimental & Applied A carology , 1 (1985) 35--44 35

Elsevier Science Publishers B.V., Amsterdam -- Printed in The Netherlands

A CARBON DIOXIDE TRAP FOR P R O L O N G E D S A M P L I N G OF

I X O D E S R I C I N U S L. P O P U L A T I O N S

J.S. GRAY
Department o f Agricultural Zoology and Genetics, University College, Belfield, Dublin 4
(Ireland)
(Accepted 20 December 1984)

ABSTRACT

Gray, J.S., 1985. A carbon dioxide trap for prolonged sampling of Ixodes ricinus L.
populations. Exp. Appl. Acarol., 1: 35--44.

A carbon dioxide trap designed to capture Ixodes ricinus over periods of up to 7 days
is described. The trap compared favourably with blanket dragging and flagging in areas of
high tick density, particularly on rough ground and for adult ticks; it was also surprising-
ly efficient for larvae. Ticks appeared to be captured throughout the 7-day trapping
period. The maximum attraction distance recorded for adult female ticks was 3.5 m and
for nymphs 1.0 m. Trapping rates were influenced by air temperature.

INTRODUCTION

Active p o p u l a t i o n s o f I x o d e s ricinus have b e e n sampled b y a variety o f

m e t h o d s , m o s t of w h i c h m a y be allocated to o n e o f t w o categories. T h e first
o f these categories involves c o l l e c t i o n o f ticks over a p e r i o d o f days during
which the investigator has little c o n t r o l o f t h e actual pick-up o f the ticks.
Such m e t h o d s include the use o f t r a c e r animals (eg. sheep o r cattle) (Milne,
1943; Barnard, 1981), and the use o f t r a p p e d r o d e n t s f o r sampling the
i m m a t u r e stages ( M e r m o d et al., 1973; Campbell and MacKay, 1979). B o t h
these m e t h o d s have the advantage o f being realistic in t e r m s o f tick pick-up
and in being long-term sampling regimes. T h e y suffer in t e r m s o f practicality
and e x p e n s e and are m o s t l y used in intensive localised studies.
T h e s e c o n d c a t e g o r y are relatively s h o r t - t e r m m e t h o d s consisting o f
the use o f flags or blankets w h i c h are b r o u g h t into c o n t a c t w i t h questing
ticks (Philip, 1937; Milne, 1943). A l t h o u g h t h e investigator has a g o o d deal
o f c o n t r o l o f the sampling p r o c e d u r e in these m e t h o d s , t h e r e is a disad-
vantage in t h a t the sampling is carried o u t over a s h o r t period o f t i m e so
t h a t , w h e n w e a t h e r c o n d i t i o n s and vegetational s t r u c t u r e vary, misleading
results m a y be o b t a i n e d . T h e y have the f u r t h e r disadvantage in t h a t t h e y are
inefficient for collecting adult ticks ( S u t h e r s t et al., 1 9 7 8 ) .

0168-8162/85/$03.30 9 1985 Elsevier Science Publishers B.V.

36

A third approach to sampling consists of attracting ticks to carbon dioxide

traps. This method overcomes problems associated with the nature of the
vegetation and also seems to be particularly suitable for sampling adult ticks.
Such traps are usually designed to operate for only a matter of a few hours,
but in two studies trapping continued for 24 h (Wilson et al., 1972) and 72 h
(Eads et al., 1982). In the present study an attempt was made to overcome
the problem of short sampling time to some extent by using a trap designed
to attract and trap I. ricinus for up to 7 days.

MATERIALS AND METHODS

Trap design

The design of the trap is based on that described by Wilson et al. (1972),
but enlarged to accommodate more dry ice and thus to continue trapping for
a longer period. The trap (Fig. 1) is made of fibreglass foam with a lining in-
side and out of solid fibreglass, and consists of a box (A) with an internal
dimension just large enough to accommodate a 12 kg block of dry ice. The
lid (B) is made of thick fibreglass foam, and is designed to fit into the top of
the box. Draught excluder lines the rim of the box and the lid is clamped

Fig, 1. Carbon dioxide trap for the capture of I x o d e s ricinus. (A) Dry ice box; (B) lid; (C)
base platform; (D) platform roof; (E) sticky tape; (F) diffusion apertures. All measure-
ments in cm.
 37

down onto it with two bolts and butterfly nuts. The platform (C), also of
fibreglass, is not attached to the box but surrounds it just below the level of
3 mm diameter diffusion holes (F), one of which was drilled in each side of
the box at an upward angle of about 30 ~, into the b o t t o m of the carbon
dioxide compartment. The platform forms an angle with the ground of
about 20 ~ A second similar platform (D) is positioned around the trap about
10 cm above the first as weather protection. The whole trap weighs 6.5 kg.
To activate the trap, a single 12 kg block of dry ice is placed in the box.
The CO2 diffuses through the holes in the box wall and disperses over the
area surrounding the trap. Ticks attracted to the trap walk up the sloping
sides of the platform and are trapped by sticky tape (E), which is 5 cm wide
and placed sticky side down on the upper rim of the platform, so that half
of the tape overhangs the sloping sides. It was found t h a t ordinary masking
tape, which was used by Wilson et al. (1972), did not perform well for more
than 2 days so stickier, water resistant carpet tape (Warrior Products Limit-
ed, London) was used.

S t u d y area

The traps were used in hill land in Co. Wicklow at two separate sites. Site
1 was an unfenced area of approximately 50 acres in a valley bottom to
which sheep and cattle had access throughout the year. Site 2 was an area of
about 70 acres divided into small fields, and which was unrestrictedly grazed
by a herd of six cows and their calves from May to December each year. The
studies were carried out in early summer (May to early July) at both sites,
at which time the vegetation consisted predominantly of coarse grasses
(Agrostis spp.), dead bracken fern (Pteridium aquilinum) and fern shoots up
to about 350 cm in height.
No specific study was carried out to compare tick densities at the two
sites but it was noted that during peak nymphal activity at Site 1 an average
of approximately 60 nymphs were captured by a 25 m blanket drag com-
pared with an average of 30 at Site 2.

Sa m p ling p roced u re

A single CO2 trap was used at Site 1 (Study I, 1978) and was replenished
with a 12 kg block of dry ice each week from early May to mid-July for a
total of nine trappings. Three traps were used at Site 2 (Study 2, 1982) and
the traps were operated every second week instead of weekly because of dry
ice supply problems. Samplings were made from late May to late August for
a total of eight trappings.
At Site 1 blanket-drag samplings were also carried out and this consisted
of dragging a 1.75 • 1.15 m blanket for 25 m, turning it over, dragging it a
further 25 m, and then placing it in a plastic bag for subsequent examination
in the laboratory. Five such samplings were made once a week between 1200
and 1400 h in an area adjacent to the trap.
38

At Site 2 blanket dragging was replaced by the strip-flag method. This

method was developed after Study 1 was completed (Gray and Lohan,
1982) and is considered to be superior to blanket-drag sampling for use on
rough ground. The strip-flag consists of a synthetic fibre white blanket cut to
1.0 X 0.7 m and then cut along its length into ten 65-cm strips. The flag was
attached to a 1.75 m batten and swept in a semicircle in front of the operator.
Each sampling consisted of ten sweeps with two paces between each sweep.
Ten samplings were carried out each week between 1100 and 1400 h from
early May to late August.

Mark and recapture studies

During Study 2 (June 1982) a total o f 350 male, 350 female and 300
nymphal unfed wild ticks were captured by flagging, marked with cellulose
paint and deposited at measured distances (0.5, 1.0~1.5, 2.0, 3.0, 3.5 and 4.0
m) from the traps. The capture rates were assessed 7 days later. This was
carried out on three occasions using three traps for the first two occasions
and four for the last.

Me te o ro logy

Weather data were obtained from thermohygrographs in Stevenson screens

situated at each site. Temperature readings were also taken at ground level at
the time of blanket and strip flag sampling.

RESULTS

Study 1 (Site 1, 1979)

The total numbers of ticks captured by the CO2 trap were 4777 larvae,
1092 nymphs, 453 females and 319 males and by blanket dragging 299 lar-
vae, 1322 nymphs, 100 females and 99 males.
It is evident that the CO~ trap was more efficient than the blanket drag
for both adults and larvae during their normal peak activity periods (May
and July, respectively), whereas at least as m a n y n y m p h s were captured
with the blanket as with the trap.
The patterns of activity determined by the trap for larvae, nymphs and
adults were remarkably similar (Fig. 2), despite the fact that the three stages
could be expected to have different distributions as a result of differing
seasonal activity and engorgement periods. The results for the different
stages of ticks captured by blanket dragging did not correlate nearly as well.
The drag method does not seem to have been at all suitable for larvae in this
particular habitat.
The number of ticks of all three stages captured by traps were obviously
influenced by air temperature (Fig. 2), though statistical significance as de-
 39

CO2 Trap Blanket-drag

22
T ~ 20

g ~16~

12
lq
|

,<
~ \\\\\

Larvae

r7 rn fl n -_~- I-1__15
3-
Nymphs

nl NRnn
2
2-

Z 3- Dg Adults

. . . .
M J J M J J
Fig. 2. C o m p a r i s o n o f tick c a p t u r e b y a single c a r b o n d i o x i d e t r a p a n d b y b l a n k e t dragging;
air t e m p e r a t u r e s at sampling. S t u d y 1, M a y - - J u l y 1 9 7 8 , Co. Wicklow, Ireland.

140-
J
7
7

./

/ ,7
100- ./.f. / 7"7

.u
i-

60-
2

E I /
Z / ~

20-

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Days after trap activation

Fig. 3. C u m u l a t e d t o t a l n u m b e r s o f I. ricinus c a p t u r e d b y c a r b o n d i o x i d e t r a p over 7
days. S t u d y 1, Site 1, M a y 1 9 7 8 . (-- 9 --) A d u l t s ; (--) n y m p h s ; (- - -) larvae.
40

termined from correlation coefficients was not obtained because of the limit-
ed number of observations. No such relationship was observed for the ticks
captured by blanket dragging.
It would seem that ticks continued to reach the sticky tape on the trap for
at least 7 days after activation during peak adult activity (Fig. 3) and for at
least 5 days during peak larval activity (Fig. 4). The rate of capture declined
in the second half of the trapping period in most cases, but it is probable
that ticks of all stages would continue to be trapped beyond this period if
more CO2 was provided.

10-
I
/
/
/
/
/
/
/
/
8- /
/
/
I
/
/
i
/
6- /
/
/
ks
1
/
/
"6 4- /
/
/
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_o /
E
D
z
2-

iIII"z
1 2 ~ 4 ;
Days after trap activation

Fig. 4. Cumulated total numbers of I. ricinus captured by carbon dioxide trap over 5
days. Study 1, Site 1, July 1978. ( - - , - - ) A d u l t s ; (--) n y m p h s ; (- - -) larvae.
 41

Study 2 (Site 2, 1982)

In this study the CO2 traps seemed to have performed adequately in an

area of lower tick density (Fig. 5). Good agreement in the pattern of results
was obtained with the two sampling methods for larvae and nymphs but not
for adults, for which the CO2 traps seemed to be much more efficient as
judged by the detection of the expected spring peak. The total numbers of
ticks captured by the CO2 traps were 1940 larvae, 1331 nymphs, 100 females
and 72 males and by the strip-flags 8333 larvae, 2269 nymphs, 35 females
and 57 males. As in Study 1, air temperatures were found to influence
trapping rates, though not as strongly. The different number of replicates
CO2 Trap Strip- Flag
u 22.]
Er 20 ...
>.~

24 Larvae

n R
u
Nymphs

D~ Adults
Z 14-~
~.o- ! [~ 0[] r~~r
0<5-

0-2- ~ 4] rT/f r ~ ~ ~, o,.~ ~,

M J J f A M J J A
Fig. 5. Comparison of tick capture by three carbon dioxide traps and by strip-flagging; air
temperatures at sampling. Study 2, Site 2, May--August 1982, Co. Wicklow, Ireland.

TABLE i

Data variability from CO 2 traps and strip-flag sampling methods as assessed from average
variance to mean ratios (Study 2)

Sample methods Larvae Nymphs Females Males

CO~ trap
(three replicates, eight observations) 146.5 10.7 3.5 0.71
Strip-flag
(ten replicates, eight observations) 5.2 3.2 1.1 0.6
42

used in the two m e t h o d s make it difficult t o c o m p a r e variability in the data;

however, average variance t o m e a n ratios indicate t h a t in this respect the
CO2 trap d a t a f o r n y m p h s and adults c o m p a r e d r e a s o n a b l y well w i t h the
strip-flag data (Table 1). T h e great variability s h o w n in t h e CO2 t r a p data for
the larvae reflects b o t h t h e highly aggregated d i s t r i b u t i o n and the low
m o b i l i t y o f this stage.
TABLE 2

Percentage marked ticks recaptured by CO 2 trap (Study 2, Site 2, May--June 1982)

Tick No. of No. per Distance deposited from trap (m)

stage traps deposition
used 0.5 1.0 1.5 2.0 3.0 3.5 4.0

Nymph 3 100 -- 9 0 0 --
Female 7 50 32 12 32 8 16 2 0
Male 7 50 24 8 12 6 2 0 0

T h e mark and r e c a p t u r e s t u d y s h o w e d t h a t m o s t ticks failed to l o c a t e the

traps even w h e n d e p o s i t e d as close as 0.5 m, b u t a few individuals r e a c h e d
the traps f r o m m u c h greater distances (Table 2). N y m p h s were n o t a t t r a c t e d
f r o m distances o f m o r e t h a n 1 m whereas adults were a t t r a c t e d f r o m u p t o
3.5 m. No a t t e m p t was m a d e to measure larval m i g r a t i o n because o f diffi-
culties in marking larvae.

DISCUSSION

Several a u t h o r s (Milne, 1943; S u t h e r s t et al., 1978) have stated t h a t

b l a n k e t dragging or flagging does n o t result in g o o d c a p t u r e rates of adult
ticks due to their failure t o attach, and also t c the fact t h a t their large size
means t h a t t h e y are m o r e readily k n o c k e d o f f a f t e r a t t a c h m e n t t h a n are
larvae and n y m p h s . Since the CO2 trap relies on attracting ticks to it for
capture, it is not surprising t h a t adult ticks, t h e m o s t m o b i l e b u t least
n u m e r o u s o f the t h r e e stages, were c a p t u r e d in relatively large numbers. The
sex ratio, female : male, was 1.45 : 1.00 for the CO2 trap c o m p a r e d w i t h
0.98 : 1.00 f o r t h e b l a n k e t samplings in S t u d y 1 and 1.44 : 1.00 (CO2 trap)
and 0.70 : 1.00 (strip-flag) in S t u d y 2. This d i f f e r e n c e is explicable in t e r m s
o f the greater m o b i l i t y o f the larger female and also the greater t e n d e n c y
o f the female to be k n o c k e d o f f t h e b l a n k e t and strip flag. It should be
n o t e d t h a t the m o s t efficient flagging m e t h o d for adult c a p t u r e was n o t
used here. This consists o f inspection o f an entire flag and removal o f ticks
at every sweep f o r a ten m i n u t e sampling period. Its main d r a w b a c k and t h e
reason it was n o t used here is t h a t i m m a t u r e stages c a n n o t be sampled at the
same time as it w o u l d be t o o time c o n s u m i n g to r e m o v e t h e m at each sweep.
T h e n u m b e r s o f n y m p h s caught b y the m e t h o d s used in b o t h S t u d y 1 and
2 are m o r e c o m p a r a b l e t h a n for adults m a i n l y because b l a n k e t dragging/
 43

flagging is more efficient for nymphs and compares reasonably well with the
use of tracer sheep (Gray et al., 1978).
The results obtained in Study 1 for the larvae showed that the CO2 trap
was surprisingly efficient, and that the blanket trap appeared much less so.
Although blanket dragging has proved an adequate sampling m e t h o d on uni-
form ground (Gray et al., 1978), the ground used in these studies was quite
rough. It is probable that many larvae do not attempt to reach the vegeta-
tion surface of the vegetation where they would contact the blanket (Gray,
1981). This tendency would enhance the relative efficiency of the CO2 trap.
The strip flag, used in Study 2, was designed for larvae and the results
produced were much more comparable with those obtained with the CO2
trap (Fig. 5). Nevertheless, the CO2 trap still seemed to be more sensitive and
detected larvae earlier and at a slightly higher level.
The similarity of the patterns of activity for the ticks caught by the CO2
trap suggests that the response to CO2 by the three stages is consistent and
also that the trap catchment area is sufficiently large to overcome the aggre-
gated distribution of the ticks in areas of high tick density. The much smaller
numbers of ticks captured at Site 2 compared with Site 1 is only partly due
to lower tick density. The vegetation was considerably thicker at Site 2 and
it is probable that this factor influences capture rates.
The distances that I. ricinus was found to migrate to the CO2 trap were
much smaller than those found for A m b l y o m m a americanum (Wilson et al.,
1972) and D e r m a c e n t o r andersoni (Eads et al., 1982), b u t were surprisingly
large in view of the observations of Milne (1950) on horizontal movement of
unfed I. ricinus. In fact it is probable that this species is capable of greater
movement than the 3.5 m maximum found in the present study, since the
capture and marking of wild ticks probably affects their viability.
It would seem that the population of I. ricinus available to the CO2 trap is
not exhausted even after seven days. Wilson et al. (1972) found that 20--
40% of A. americanum attracted to the trap within 24 h did not climb onto
the trap but ascended vegetation in the immediate vicinity. It is not k n o w n
whether this applies to I. ricinus as well, b u t it could explain the continued
trapping of I. ricinus over the 7-day test period.
It is not surprising that air temperatures have such a marked effect on tick
capture rates by CO2 traps, since the ticks must be active to migrate to the
trap and this is dependent on air temperature. In contrast, air temperatures
above the threshold for tick activity (approx. 7~ were not found to in-
fluence capture rates b y blanket dragging or flagging, which confirms
previous observations (Gray et al., 1978; Gray, 1980). It has also been ob-
served that, once air temperature reaches this activity threshold, it has a
limited effect on pick-up rate by tracer sheep (Gray et al., 1978). Therefore
it is likely that samples from CO2 traps are slightly biased under Irish condi-
tions, where most ticks are probably picked up b y moving hosts at subop-
timal temperatures for activity.
It is evident from these studies that the 7-day CO2 trap can successfully
44

overcome problems associated with short sampling times, but its usefulness
is probably limited to areas of high tick density.

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Barnard, D.R,, 1981. Comparison of populations of ticks free living on pasture and
parasitic on cattle. Ann. Entomol. Soc. Am., 74: 507--511.
Campbell, A. and MacKay, P.R., 1979. Distribution of the American dog tick, Dermacen-
tor variabilis (Say) and its small-mammal host in relation to vegetation in a study area
in Nova Scotia. Can. J. Zool., 57: 1950--1959.
Eads, R.B., Smith, G.C. and Maupin, G.O., 1982. A CO: platform trap for taking adult
D e r m a c e n t o r andersoni (Acari: Ixodidae). Proc. Entomol. Soc. Wash., 84: 342--348.
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stages under field conditions. Bull. Entomol. Res., 71: 533--542.
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Appl. Biol., 30: 240--250.
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the adult tick. Parasitology, 40: 14--34.
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Sutherst, R.W., Wharton, R.H. and Utech, K.B., 1978. Guide to studies on tick ecology.
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Lone Star tick (Acarina: Ixodidae). 1. Response of different developmental stages to
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