Journal of Veterinary Parasitology 11(2) 1997: 155-159

IN VITRO EFFICACY OF HERBAL FORMULATION AV/EPP/14* AGAINST DOG TICK

RHIPICEPHALUS SANGUINEUS
D.N. Panda and S.C. Misra
Department of Parasitology, College of Veterinary Science & Animal Husbandry, OUAT,
Bhubaneswar - 751 003, India

ABSTRACT
In vitro efficacy of the herbal formulation AY/EPP/14 at 1:0, 1:1, 1:2, 1:3, 1:4, 1:5 and 1:6 concentrations
was evaluated against the common dog tick, Rhipicephalus sanguineus by exposing batches of ticks for 15,
30, 45 and 60 minutes in each concentration. There was 100 percent mortality of the nymphal stages
treated with 1:0 to 1:4 concentrations. At 1:5 and 1:6 dilutions the mortality rate varied from 93-100
percent and 82-98 percent, respectively. Against adult females, the drug resulted in 100 percent mortality at
1:0 and 1:1 concentrations. At concentration of 1:2, 1:3, 1:4, 1:5 and 1:6, the percent mortality ranged
between 88-98, 70-92, 47-83, 28-60 and 20-50, respectively according to the length of exposure. The
percent reduction in egg laying of surviving females was 94-100, 76-95, 55-89, 35-72 and 27-64,
respectively. There was only slight adverse effect on the hatchability of eggs. The mortality rate of
nymphs and adults, and fecundity of females had a positive correlation with the exposure time.
INTRODUCTION
Control of dog ticks is mainly done by intensive and continuous application o/ synthetic, non-
biodegradable acaricides on the host as well as its surroundings. But the acaricidal hazards like
toxicity to animals and human beings coupled with immunosuppression. environmental pollution,
and development of acaricidal resistance (Nolan, 1987) has necessitated search for safer, environment-
friendly, novel plant derived acaricides. The in vitro efficacy of one such herbal product, AV/EPP/14,
against Rhipicephalus sanguineus is reported here.
MATERIALS AND METHODS
Engorged nymphs and adult females of R. sanguineus were collected in sufficient numbers from
naturally infested dogs. The test procedure of Khan and Srivastava (1977) with modifications regarding
time of dipping was followed (Sinha et al., 1987}.Batches of 20 nymphs and 20 females were placed
separately in between the folds of filter papers soaked in 7 different freshly prepared dilutions of
AV/EPP/14 in tap water viz., 1:0,1:1, 1:2, 1:3, 1:4, 1:5 and 1:6. In each concentration, the ticks were
retained for four different time durations, viz., 15, 30, 45 and 60 minutes. Concurrently, control
batches were treated with tap water on similar lines.
After treatment, the ticks of both treated and control groups were transferred to petridishes,
examined with a hand lens and the ticks showing no progressive movement were counted as dead.
The remaining ticks were incubated at 28±2°C and 85±5 percent R.H. for 48 hr and the mortality
pattern at 4, 8. 12, 24 and 48 hr was recorded. The engorged females that survived 48 hr after
treatment were further incubated to observe the effect of the drug on egg laying and hatching.
RESULTS
The acaricidal property of the drug was observed to vary in direct proportion to the drug
concentration and exposure time. Further, the immediate, subsequent and cumulative killing effect
of the drug was more pronounced for nymphs than adult females (Table 1 and 2). As such, it was
found 100 percent effective against nymphal stages up to 1:4 dilutions and against adults up to 1:1
dilution. The efficacy against adults varied from 88-98, 70-92, 47-83, 28-60 and 20-50 percent at
different exposure times, in 1:2, 1:3, 1:4, 1:5 and 1:6 dilutions, respectively.
The engorged females of control group on an average laid 3668 eggs with a natural viability
of 93.7 percent, whereas, 100 percent mortality occurred in female ticks after treatment with 1:0 and
1:1 dilution of the drug. In the females treated with 1:2, 1:3 and 1:4 dilutions, there was substantial
reduction in egg laying ranging between 94-100, 76-95 and 55-89 percent, respectively. The egg
hatchability appeared only marginally lowered in inverse proportion to exposure time (maximum
observed reduction 12 percent in 1:2 dilution, 45 minutes exposure time).
DISCUSSION
In order to reduce reliance on chemical acaricides, there is a need to diversify and increase
the choice of non-toxic alternatives after their systematic evaluation both in vitro and in vivo. The
efficacy of herbal formulations against mange and other ectoparasites in dogs has earlier been
reported (Das, 1996; Mignon and Losson, 1996; Sharma, 1996). The herbal ingredients of AV/EPP/14
viz., Acorns calamus (Deshmukh et al., 1982), Azadirachta indica (Singh, 1983) Pongamia glabra (Kale
and Pachgaonkar, 1969), Cedrus deodara (Lai et al., 1976) and Eucalyptus globulus (Chavan et al.,
1982) are known for their insecticidal, growth inhibitor)', anti-moulting, anti-fertility and repellent activities. In
the present study, AV/EPP/14, apart from marked lethal effects on fed nymphs, was found 100
percent effective against adult R. sanguineous in single treatment of 1:1 dilution. There was heavy
mortality with the use of 1:2, 1:3 and 1:4 dilutions
Table 1. In vitro efficacy of AV/EPP/14 against Rhipicephalus sanguineus

No. of
ticks No. of ticks died , times in Minute
Dilution treated
per time 15 30 45 60
duration

Treated A N A N A N A N A N
1:0 60 60 16(27) 50(83) 23(38) 60(100) 31(52) 60(100) 37(62) 60(100)
1:1 60 60 11(18) 35(58) 18(30) 51(85) 23(38) 58(97) 27(45) 60(100)
1:2 60 60 7 (12) 26 43) 14(23) 40(67) 19(32) 49(82) 24(40) 58(97)
1:3 60 60 8 (13) 20(33) 13(22) 38(63) 20(33) 45(75) 22(37) 51(85)
1:4 60 60 5 (8) 22(37) 10(17) 33(55) 16(27) 42(70) 18(30) 48(80)
1:5 60 60 2 (3) 19(32) 7(12) 26(43) 10(17) 33(55) 13(22) 39(65)
1:6 60 60 -- (0) 14(23) 3 (5) 18(30) 7(12) 27(45) 11(18) 31(52)
CONTROL

Tap 60 60 60 60 60 60 60 60 60 60
water (0) (0) (0) (0) (0) (0) (0) (0) (0) (0)
N = Nymphs A = Adult females (engorged)

However, there lies the need for repeating the treatment for complete control. In addition, the
average egg laying of the surviving females was drastically reduced thereby adding another dimension
to overall control. The continuous mortality observed till 48 hr post-treatment indicates that the
subsequent application can be made after an interval of atleast48 hr. The relatively much longer
exposure time required for herbal ectoparasiticides (Sinha et al., 1987) compared to chemical agents
may be viewed from the aspect that plant based acaricides are a separate class in themselves.
Moreover, the prolonged contact with the formulation neither causes discomfort nor there is toxicity
risk unlike chemical acaricides (Sharma, 1996). Although, the action of AV/EPP/14 is somewhat
delayed as compared to synthetic pesticides, the ultimate effect in killing or inhibiting the ticks was
almost identical. However, there is need to further substantiate the present observations through in
vivo trials. lity pattern of ticks after incubation
Table 2.Invitro efficacy of AV/EPP/14 against R.sanguineus : Morta

No. of ticks Max.

No. of ticks
incubated(N) per Max. Mortality in mortality in
incubated(A) per
Concentration each exposure time of incubation time of
each exposure
time (%) incubation
time
(%)
15 30 45 60 15 30 45 60
All died
All died within 4 hrs
1:0 10 - - - 44 37 29 23 within 8-
(100)
24hr (100)
All died
All died within 4-8
1:1 25 9 2 - 49 42 37 33 within 12-
hrs (100)
48hr (100)
Partial
All died within 4-24
1:2 34 20 11 2 53 46 41 36 (cumulative
hrs (100)
88.3-98.3)
Partial
All died within 8-24
1:3 40 22 15 9 52 47 40 38 (cumulative
hrs (100)
70.0-91.7)
Partial
All died within12-48
1:4 38 27 18 12 55 50 44 42 (cumulative
hrs (100)
46.7-83.3)
Of 60min exposure all
Partial
died in 24 hrs others
1:5 41 34 27 21 58 53 50 47 (cumulative
substantially till 48 hr
28.3-60.0)
(93.3-100)
Partial in all till Partial
1:6 46 42 33 29 (81.7-98.3) 60 57 53 49 (cumulative
48 hr. 20.0-50.0)
N- Nymphs A- Adult females (Engorged)

Research code name of new ectoparasiucide formulation for pets. Dabur Ayurvet Limited, 22 Site IV,
Sahibabad, Ghaziabad - 201 010