Tax►ticimy

cit African Ticks

An Identification Manual

Jo5eph Okello-Onen, 5hawgi f I Ha55an

and 5uliman E55Urridil

partageons les Lkirtnawukteas au prnf1 dits ccirnmunautis ruraJes

sharing knowledge, Improving rural livatthocids

II
TAXONOMY OF AFRICAN TICKS
An Identification Manual

Joseph Okello-Onen, Shawgi M. Hassan

and Sunman Essuman
 TAXONOMY OF AFRICAN TICKS
An Identification Manual

Joseph Okello-Onen', Shawgi NI. Hassan' and Suliman Essutnan2

'Livestock Health Research Institute (LIM). P.O. Box 96, Toraro, Uganda

International Centre of Insect Physiology and Ecology (ICIPE)

P. 0. Box 30772-00100 Nairobi, Kenya

Edited by Serah W. Mwanycky and Annalee Ngieny Mengech -

Illustrations by N. Mwanga Komori

International Centre of Insect Physiology and Ecology (ICIPE)

African Regional Postgniduate Programme in Insect Science (ARPP1S)
P.C1 Box 30772-00100 Nairobi. Kenya.
Tel: +254 (20) 8632000; Fax; +254 (20) 8632001/2
E-mail: icipee'icipc.org
Special Acknowledgement
ICIPE gratefully acknowledges the Technical Centre for Agricultural and Rural
Cooperation (CTA) for their sponsorship ()Effie reprinting of this manual.

The Technical Centre for Agricultural and Rural Cooperation (CTA) was establ ished in
1083 under the Lome Convention between the ACP (African, Caribbean and Pacific)
Group of States and the European Union Member States. Since 2000_ it has operated
within the framework oldie ACP-EC Cotonou Agreement.

CTA's tasks are to develop and provide services that improve access to information for
agricultural and rural development. and to strengthen the capacity of ACP countries to
produce. acquire, exchange and utilise information in this area_ CTA's programmes arc
designed to; provide a wide range of information products and services and enhance
awareness of relevant information sources; promote the integrated use of appropriate
communication channels and intensify contacts and information exchange (particularly
intra-ACP): and &Niel op AC P capacity to generate and manage agricultural information
and to formulate ICM strategies, including those relevant to science and technology.
CTA's work incorporates new developments in methodologies and cross-cutting issues
such as gender and social capital,

CTA
Postbus 380
6700 A.1 Wageningen
The Netherlands

Website: www-cta.int

Published arid printed by [OPE Science Press

P.0, Box 30772-00100 Nairobi, Kenya
Tel: +254 (20) 8632000
Fax; +254 (20) 863200112

01999 The International Centre of Insect Physiology and Ecology (ICIPE)

First Edition 1999. First impression 2006

All rights reserved. No part of this publication may be reproduced in any

form without the written permission of ICIPE ICIPE Science Press
—

Citation: Okeflo-Onen J., Hassan S. M. and Essu in an S. (1999)

Taxonomy of African licks: An Identification Manual. 124 pp.
ICIPE Science Press, Nairobi
ISBN: 92 9064 127 4
October 1999
Reprinted January 2006
CONTENTS
Page

List of Illustrations and Tables vi

Foreword ix

Acknowledgements xi

1.0 General Introduction 1

2.0 Methods of Collecting Ticks 3

2.1. Restraining of Hosts 3
2.1.1 Domestic Animals 3
2.1.2 Wild Animals 3
2.2 Collection of Ticks 3
2.2.1 Sampling Criteria 3
2.2.2 Collecting Parasitic Ticks 5
2,2.3 Collecting Non-Parasitic Ticks (Host-Seeking) 5
2.2.3.1 Blanket Dragging Method 7
2.2.3.2 Flagging Method 7
2.2.3.3 Carbon Dioxide Trap 7
2.2.3.4 Pheromone Trap
2.3 Preservation and Labelling of Ticks 9
3.0 Rearing of Ticks for Identilication 10

4,0 Requirements For Tick Taxonomy 12

5.0 Introduction to Identification of Ticks 13

5.1 Differentiation of Major Groups of Ticks 13
5.1.1 Differences Between Argasidae (Soft) and
Ixodidae (Hard) Ticks 14
5.1.2 Genera of Ticks 14
5.1,3 Differences Between Male and Female Ticks 23
5.1.3.1 Argasidae (Soft ticks) 23
5.1.3.2 Ixodklae (Hard ticks)
5.1.4 Differences Between Adult and Immature Ticks 23
6.0 Identification of Ticks of Economic Importance 26
6.1 Family Argasidae 26
6.1.1 Genus Arga.i 26
6.1.2 Genus Ornirhodorns 26
6.1 Family lxodidae 32
6.2.1 Genus Amblyommo 32
6,2.2 Genus Boophllus 42
6.2.3 Genus Hyaininma 67
6.2.4 Genus Rhipicephalus 87

7.0 Important References 123

Annex 1: A Rapid Method for Clearing and Mounting the
Genitalia or Female Ixodid Ticks Al
 LIST OF ILLUSTRATIONS AND TABLES

FIGURES

FIGURE 1 Method of restraining an animal before casting on ground; steps 1-4. 4

The predilection sites (attachment sites) of major tick species 6
FIGURE 2
Metliods of collecting host-seeking ticks in pasture 8
FIGURE 3
Genus Argas: Key morphological characters used in identification 17
FIGURE 4
FIGURE 5 Genus Oruithudoros: Key morphological characters used in identification 18
FIGURE 6 Genus Lvodes (male and female): Key identification features on dorsal
19
and ventral sides
Key morphological characters used in identification of ixodid ticks 20
FIGURE 7
FIGURE 8 Important morphological characters for distinguishing the species of
Boophilus ticks 57
FIGURE 9 Important morphological characters for distinguishing the species of 92
Rhipicephalus ticks

FLOW CHARTS
FLOW CHART 1 Key distinguishing morphological characters for tick genera in the
family Ixodidae 21
FLOW CHART 2a Genus kith/walnut: Key distinguishing morphological characters (Males) 36
FLOW CHART 2h Genus Andaiye►r►►ti: Key distinguishing morphological characters (Females) 37
FLOW CHART 3 Genus Boophilus : Key distinguishing morphological characters 44
FLOW CHART 4a Genus Hyalomma: Key distinguishing morphological characters (Males) 71
FLOW CHART 4b Genus Hyalomma: Key distinguishing morphological characters (Females) 74
FLOW CHART 5a Genus Rhipicephalus: Key distinguishing morphologidal characters (Males) 93
FLOW CHART 5b Genus Rhipicephohts: Key distinguishing morphological characters (Females) 96
FLOW CHARY 5c
Genus Rhipicephalus: Ornamented group (females) 99

PLATES (Electron micrographs)

PLATE I Larva and nymph of Rhipicephaias appendiculatus
PLATE 2 Argas persicus (male) 25
PLATE 3 Omithodoms moithato (male) 29
PLATE 17 Boophilus a►miatus (male and female) 31
PLATE 18 Boophilus decohn-atus (male and female) 59
PLATE 19 Boophilus geigyi (female) 61
PLATE 20 Boophilus mieroplus (male and female) 63
PLATE 21 Hyolomma anatolicum anatolicum (male and female) 65
PLATE. 22 Hy/tont/no dromedarii (male) 77
PLATE 23 Ilyalomma hupehaium (male and female) 79
PLATE 24 Hyalomma marginatum Wipes (male and female) 81
PLATE 25 Hyalomma irunceuum (male and female) 83
PLATE 26 Rhipicephalus appendlculams (male and female) 85
PLATE 27 Rhipicephalus zainbcziensis (male and female) 102
PLATE 28 Rhipicephalus evertsi evertsi (male and female) 103
PLATE 29 Rhipicephalus pfilchellus (male and female) 105
PLATE 30 Rhipicephalus sanguineus (male and female) 108
PLATE 31 Rhipicephalus shims (male and female) 111
115

vi
 COLOUR PLATES

PLATE 4 Genus Argil s (female): Key identification features on dorsal and ventral sides 45
PLATE 5 Genus Ornithodoros (male and female): Key identification features on
45
dorsal and ventral sides
PLATE 6 Genus An/boy/ma (male and female): Key identification features on 46
dorsal and ventral sides
PLATE 7 Genus Apononuna (male): Key identification features on dorsal and 47
ventral sides
PLATE 8 Genus Boop►ihis (male and female): Key identification features on dorsal
and ventral sides 49
PLATE 9 Genus Lkrinacentor (male and female): Key identification features on
50
dorsal and ventral sides
PLATE 10 51
Genus Haemaphysafis (male and female): Key identification features
on dorsal and ventral sides
PLATE I I 52
Genus F-Iyalonnna (male and female): Key identification features on
53
PLATE 12 dorsal and ventral sides
Genus Rhipieephalus (male and female): Key identification features 54
on dorsal and ventral sides
PLATE 13 Key morphological characters for And)lyonuna ticks (male and female) 55
PI ATE 14 The patterns of ornamentation on Ambiyomma ticks: (1) A. cohaerens 56
(male and female); (2) A. lepid►on (male and female)
PLATE 15 The patterns of ornamentation on Anzblyomma ticks: (1)A. gennna
(male and female); (2) A, hebraeum (male and female)
PLATE 16 The patterns of ornamentation on Amblyom►a variegation (male and female)

TABLES

TABLE I Basic morphological features used for distinguishing between soft and hard ticks 15
TABLE 2 The genera and number of tick species in the family Ixodidae 16
TABLE 3 The distinguishing morphological features between adult and immature ticks
TABLE 4 Genera Argus anti Ornithadoros: The predilection sites, distribution, host and
disease relationships 27
TABLE 5 Key morphological characters for identifying the soft tick Aix persicus 98
TABLE 6 Key morphological characters for identifying the soft tick Ornithodorox
nioubata complex 30
TABLE 7 Aniblyanona species: The predilection sites, distribution, host and disease
relationships 33
TABLE 8 Ambiyomma cohaerens: Key morphological characters 38
TABLE 9 Aniblyonona lepiehon: Key morphological characters 39
TABLE 10 An/Won/m► ►emma and A. hebraeutn: Key morphological characters 40
'TABLE 11 Amblyonona variegation: Key morphological characters
41
TABLE 12 BolThihrs species: The predilection sites, distribution, host and disease
relationships
43
TABLE 13 Boophilus annulauts: Key morphological characters 58
TABLE 14 Boophilus decolarat►s: Key morphological characters 60
TABLE 15 Boophihis geigyi: Key morphological characters 62
TABLE 16 Boophllus inicrophis: Key morphological characters 64
TABLE 17 ►orn= species: The predilection sites, distribution, host and disease
relationship
68
TABLE 18 Hyalomma amitotic-run anatolicion: Key morphological characters 76

iii
TABLE 19 Hyatt- mum, dromedarii: Key morphological character 78
TABLE 20 I-Iyalonuna impeltatum: Key morphological characters 80
TABLE 21 Hyalonuna Inarginatum rufipes-, Key morphological characters 82
TABLE 22 Hyalointna truncatu ► and H. albiparmatuni: Key morphological
characters 84
TABLE 23 Rhipicephalus species: The predilection sites, distribution, host and
disease relationships 88
TABLE 24 Groups of some rhipicephalid tick species with similar
morphological characters 91
TABLE 25 Rhipicepholus appencliculatus group: Comparative morphological
characters of closely-related tick species 100
TABLE 26 Rhipicephalus everts! everts! group: Comparative morphological
characters of males and females 104
TABLE 27 The ornate species: Comparative morphological characters of
closely-related Rhipicephalus tick species 106
TABLE 28 Rhipicephalus sanguineus group: Comparative morphological
characters of closely-related tick species. 109
TABLE 29 Rhipicephalus sinus group: Comparative morphological characters
of closely-related tick species 1I2
TABLE 30 Rhipicephalus compoSihrs group: Comparative morphological
characters of closely-related tick species 116
TABLE 31 Rhipicephalus prams group: Comparative morphological characters
of closely-related tick species 119
TABLE 32 Rhipicephalus tricuspisilunulatus group: Comparative morphological
characters of closely-related tick species I 22
Foreword

Pests and vectors arc major constraints to the improvement of crop and livestock productivity and
health conditions of man in Africa. Research on pests and vectors have been top priority areas at the
International Centre of Insect Physiology and Ecology (1CIPE), being addressed under different
research programmes, including the Ticks Research Programme.

Ticks arc reported to surpass all other arthropods in the number and variety of diseases they trans-
mit to animals and man. There are about 170 species of soft ticks (Argasidae) and 700 species of
hard ticks (Ixodidae), the majority of which are found in Africa. Several of these are known to
transmit pathogenic organisms such as protozoa, rickettsia, viruses and bacteria to animals and
man. The host and disease relationships of ticks and their distribution have been changing due to
global changes in climatic conditions, human population pressure on vegetation, unrestricted move-
ment of host animals and associations between domestic and wild animals. These trends under-
score the need for accurate tick identification to continuously re-assess the relationships between
host, vector and pathogens.

Unfortunately, the discipline of tick taxonomy has long suffered from numerous setbacks and ne-
glect. Little emphasis has been put on the significance of the discipline as a basic step in under-
standing the biology, distribution, host and disease relationships of ticks, and planning of appropri-
ate control strategies. In addition, little effort has been made to initiate young scientists into the
subject, for example through courses or workshops. As such, many scientists have had consider-
able difficulty in identifying ticks. Tick taxonomy has been monopolised by only a few devoted
scientists, mainly non-Africans. More importantly, the demand for the discipline has been weak-
ened by the dramatic effects of acaricides in controlling ticks and tick-borne diseases (TBDs).

It is not surprising that the deliberate over-dependence on acaricides for ticks and TBDs control has
diverted attention from exploring other alternative methods of control. However, it is now evident
that acaricides can no longer provide a practical, economical and sustainable solution to ticks and
TBDs control. The problems with the use of these synthetic chemicals include acaricide resistance
in ticks, pollution of the environment and food (meat and milk) with toxic residues, deleterious
effects on non-target organisms, creation of an enzootically unstable disease situation and the un-
certainty of new acaricidal molecules being produced in the near future due to prohibitive costs of
investment in research and development by the manufacturing firms. In response to the growing
impact of ticks and TBDs, especially in Africa, ICIPE has embarked on the development of alterna-
tive methods for tick control that are more specific and less toxic to non-target organisms. These
include the use of biocontrol agents, an approach which requires accurate identification of ticks if
the right enemies arc to be introduced.

tix
It is against this background that ICIPE initiated courses on tick taxonomy as a measure to strengthen
the capabilities of African scientists in the proper identification of ticks, especially those of eco-
nomic importance, so as to plan appropriate control strategies, During these courses, it was realized
that many African scientists- were not familiar with tick taxonomy and have only very limited
access to the relevant literature on the subject, especially as it pertains to African tickS. The avail-
able literature is complex and not user-friendly to a beginner, with most works being out of print or
in short supply. Most of the literature is based on work performed between 1950s and early 1970s,
and does not include the newly identified or re-described tick species.

To redress the foregoing, the production of a comprehensive manual on tick taxonomy that would
stimulate the interest of African scientists in tick unchnorny Was conceived. The task to produce this
manual was given to Drs J. Okello-Onen, S. M. Hassan and S. Essuman, who were among the
resource persons in the 1998 training course on "Novel Approaches to the Management of Ticks"
held at ICIPE Headquarters, Nairobi, Kenya.

This manual will be useful to veterinary practitioners, researchers on ticks and TBDs and univer-
sity students in Africa. It should also prove a useful resource to scientists, university lecturers and
students elsewhere. The manual was prepared with the financial support from The Netherlands
Ministry of Foreign Affairs and International Cooperation, through ICIPE's Capacity Building Pro-
gramme.

Hans Rudolf Herren, PhD Mails 0. Musewe, PhD

Di rector General, Head, Capacity Building
The International Centre of Insect Programme, ICIPE
Physiology and Ecology (ICIPE)
Acknowledgements
We are grateful to Mr M. Chi miawi of the International Centre of Insect-Physiology and Ecology
(ICIPE] for taking the electron micrographs, Mr N. Mwanga Komeri and Mr P. Lisamula for
producing the illustrations and scanning the tick photographs and to Dr Robert Copeland for taking
the tick photographs. Special thanks go to all the participants of the 1998 Training Course on
"Novel Approaches to the Management of Ticks" held at ICIPE for providing the various species of
tick specimens described in this manual. We are grateful to Drs Ellie Osir and Godwin Kaaya for
their critical review of the manuscript.

We are most grateful to Dr Vitalis 0. Musewe, Head of the Capacity Building Programme, ICIPE
for his vision in conceiving the need for a manual on tick taxonomy and to the ICIPE management
for providing us with unlimited access to all facilities needed. We thank wholeheartedly the
Netherlands Ministry of Foreign Affairs and International Cooperation, through the ICIPE Capacity
Building Programme, for their financial support.
1.0 General Introduction

Taxonomy is the discipline of science that deals with the identification of organisms, ineltiding
arthropod pests. and vectors. It is the queen of the sciences, drawing from the accumulated knowledge
of all other fields (Slater, 1960) and the starting point for all biological research (Kim, 1975). It
provides universally-acceptable scientific names to organisms and enhances meaningful scientific
communication and biological research. The discipline promotes the understanding of the
composition of the environment, the variety and interaction of all its living andnon-living components
and facilitates the development of appropriate control strategies for organisms of economic
importance.

The taxonomy of ticks is concerned with the identification of ticks belonging to two families, the
Argasidae (soft ticks) and Ixodidae (hard ticks). Ticks of economic importance to livestock in
Africa belong to the family Ixodidac. Only about 9 species (from 4 genera) out of a total of over
650 species (from 13 genera) are known to be vectors of economically important diseases. These
are Rhipicephalus (R. appendituktfus), Boophilus (B. decoloratus, B. microphi.v, B. annulatt15),
Amblyomma (A. variegatunt, A. hebraeum) and Ilyalomma (H. futotolicum anatolicum, detritunt,
H. drontedarii). They transmit a variety of tick-borne diseases (TBDs) that can be categorised into
protozoan diseases (theileriosis and babesiosis), rickettsi al disuses (anaplasmosis and cowdriosis)
and tick-associated dermatophilosis. The disease relationships of several other tick species remain
to be established. This can be achieved only if the individual tick species can he identified accurately
and their distribution and host relationships established.

Tick taxonomy, like the taxonomy of other pests and disease vectors, has often been neglected,
misunderstood or held in little regard, as evidenced by the statement, "Taxonomy is the fourth rate
game played by fourth rate scientists in fourth rate institutions" (Ride, 1978). The neglect could be
attributed to the complexity and boring nature of the subject, dominated by narrative descriptions
of diagnostic features of ticks and the use of identification keys that are, in some cases, lengthy and
confusing. This is evidenced by the fact that tick taxonomists have a tendency of using different
taxonomic criteria (e.g. different emphasis on certain morphological characters), thus creating
confusion by providing a diversity of names for certain tick species which are otherwise synonymous.

Historically, tick taxonomy has been dominated by non-African scientists, most of whom have
either died, retired or are due to retire. The only committed African tick taxonomist was the late Dr
F. P. Zulu from Zambia, who died in 1994. The discipline has not been attractive to African scientists
and little effort has been made to organise training courses in the field.

As a result of these drawbacks, the International Centre of Insect Physiology and Ecology (ICIPE)
has been organising courses on Tick Taxonomy and Tick Management with the objective of
strengthening the capabilities of African scientists in the identification and appropriate management
of ticks and TBDs. During these courses, it was realized that the relevant literature on the taxonomy
of African ticks is inadequate and unsuitable for training pioneers in the discipline. Consequently,
the resource persons for such courses have had to depend mainly on materials generated from their
personal practical experience. It is against this background that the need to produce a simple and
user-friendly manual on the taxonomy of African ticks, that would provoke the interests of scientists
in this field, was conceived. The demand for tick identification services is inevitable, especially
when implementing biological control strategies that require accurate identification of the vectors,
so that the appropriate natural enemies can be introduced. The services of tick taxonomy are also
very important in view of the widespread movement of domestic and wild animals from one place
to another, resulting in the introduction of foreign ticks, ancj extensive deforestation which causes
great changes in the microhabitat of ticks. In addition, knowledge of tick taxonomy helps in charting
out the distribution of the various species of ticks, which can then be compared with the predictions
made by the CLIMEX model.

This manual has been produced to facilitate the identification of ticks of economic importance to
livestock in Africa. It is a product of materials extracted and synthesized from books or publications
of some renowned tick taxonomists, notably 1400gstraal (1956, 1985), Yeoman and Walker (1967),
Walker (1970, 1974), Matthysse and Colbo (1987), Pegram (1984), Kaiser (1987) and De Castro
(1994). The FAO manuals on ticks and tick-borne diseases control (Volumes 1 and 2, 1984) also
provided useful information. The content of the manual has been limited to only four genera of
ticks of economic importance to livestock: Rhipicephalus, Boophilus, Amblyornma and flyalornma.
Efforts have been made to produce comprehenSive flow charts that may be more user- friendly than
the conventional identification keys, which are in some cases complicated to use The most essential
distinguishing morphological characters have been provided in tabular form for the important tick
species in the selected genera.

The identification of ticks in the genus Rhipicephalus has been simplified by first of all separating
the tick species into distinct groups based on similarities in morphological characters, then
highlighting the distinguishing morphological characters between tick species in each group. Electron
micrographs, photographs and illustrations are provided to facilitate the identification of the different
tick species.

2
2.0 Methods of Collecting Ticks
2.1 Restraining of Hosts

Ticks can he collected successfully from both domestic and wild hosts after restraining animals
using various methods. Animals are cast down for examination so that the thoroughness of
examination does not depend on the relative docility of the animals.

2.1.1 Domestic Animals

Various species of ticks infest domestic animals: cattle, goats, sheep, camels, horses, donkeys and
chickens. Ticks can be collected successfully when the animals are restrained. Cattle, horses and
donkeys can he restrained either in strong crushes or by casting down using ropes as shown in
Fig. I. The rope should be tied on the animalas shown in Steps 1 to 4, so as to avoid hurting the
animal. Goats, sheep and chickens can be restrained by physical handling. However, ticks on camels
can be collected by their owners or herdsmen.

In order to keep a good relationship with the farmers, it is advisable that the animals cast down are
not restrained for a long time. This can be made possible by having a team of persons to collect
ticks at different predilection or feeding sites as explained later.

2.1.2 Wild Animals

The large wild hosts are immobilized by anaesthesia using darts (gun or pistol). The small wild
hosts like rodents, shrews, etc. are first trapped using snap or cage traps, and later anaesthetized
under laboratory conditions.

2.2 Collection of Ticks

2.2.1 Sampling Criteria

Understanding ticks of an area or locality requires a sound sampling criterion to be developed.

Considerations should be given to host diversity, distribution, abundance and disease relationships
of different species of ticks. Whenever possible, ticks should be collected from representative samples
of both domestic and wild animals, from different eco-climatic areas. In the case of wild animals,
opportunistic collections can be made from as many animals as possible. Ticks should be collected
from domestic animals that are healthy and have not been exposed to pesticide treatment, particularly
when studying seasonal dynamics. Collections should he made from a defined or similar number of
animal species for ease of statistical analysis (De Castro; 1994).

3
 2

Fi2. 1. Method of restraining an animal before casting on ground: steps 1-4. (Adapted from Livestock
Technical Handbook, Department of Animal Production, Ministry of Agriculture, Animal
Industry and Fisheries. Entebbe, Uganda. 1995)

4
Attempts should be made to collect a reasonable number of each developmental stage of ticks
(male, female, nymphs, larvae) from different feeding or predilection sites. In case of heavy
infestations, ticks can be thoroughly collected from only half of the anima! body. Records on host
species, age, sex, health conditions, coat colour, predilection sites, etc. should also be made for
each host animal from which ticks have been collected.

2.2.2 Collecting Parasitic Ticks

Ticks infest host animals to feed for a period of about one week; the males normally stay longer on
hosts. This activity is called the "Parasitic Phase" of ticks. Different tick species prefer to feed at
different body sites of hosts called "Predilection Sites", as indicated in Fig. 2. It is, therefore,
advisable to collect ticks from these sites and keep them in separate vials or pool them together for
similar host species sampled in a locality on a particular day (morning or afternoon). This can be
done faster and efficiently by assigning a person to each-predilection site.

Ticks should be removed without causing damage, especially to the mouth parts which are useful in
identification, This can he achieved by first bending the tick on its back and then pulling it straight
upwards. The engorged females can be picked by hand or with fine curved forceps, held as close as
possible to the skin (Kaiser, 1987), The immature stages (larvae and nymphs) can be collected
using a blunt pair of forceps.

2.2 3 Collecting Non-Parasitic Tick, (Host-Seeking)

.

The non-parasitic ticks are the ticks found off the hosts in the vegetation. It comprises all the
developmental stages of ticks (larvae, nymphs and adults) both unfed and the engorged ones that
have dropped off the host to moult and lay eggs. However, it is easier to collect the unfed ticks than
the engorged ones, because of their host-seeking behaviour. These ticks quest for hosts by climbing
on vegetation, an act commonly referred to as "Ambushing". During this process. they lose some
of their body water and return to the ground when they fail to get suitable hosts or the amount of
body water lost is above the critical equilibrium humidity level. Some species of ticks (especially
Amblyonuna and Hyalomina), do not climb on vegetation, but run after the hosts once activated by
their presence. These groups of ticks are called "Hunters".

The ticks that climb on vegetation to quest for hosts can be collected by two popular methods:
blanket dragging and flagging methods.

The group of ticks which do not climb on vegetation, but hunt for their hosts (especially Amblyomma
and Hyahnouna) are collected by carbon dioxide (CO.) and pheromone traps.

5
 Ears
Shoulder/back/side
Head

Tail
Escutcheon

Dewlap/neck/brisket

Belly
Fig. 2. The predilection sites (attachment sites) of major tick species (Adapted from Livestock
Technical Handbook, Department of Animal Production, Ministry of Agriculture, Animal
Industry and Fisheries, Entebbe, Uganda, 1995)
 2.2.3.1 BLANKET DRAGGING METHOD

This method requires flannel material (1 m2) to which a wooden base is attached at one end to
stabilise it onto the ground. A piece of string is then attached at the two ends of the wooden base to
assist in pulling the flannel on vegetation, as shown in Fig. 3a. The cloth material is best dragged
at a slow walking pace, with the operator walking in front. One or two cloth materials can be
dragged by one person. Dragging is best performed in a straight line over a defined distance of
about 20 m in a transect. This activity has to be replicated so as to get unbiased density of ticks
per locality.

A fter dragging over each transect or a specified distance, the surface of the cloth material in contact
with the vegetation is folded inwards. The cloth material is spread onto a table or over a flat surface
like bonnet of a car. The ticks that attach onto the cloth material can be collected by a pair of
forceps, fine hairbrush or aspirators. Dragging method is useful for collecting the immature stages
of ticks in low to medium height vegetation,
- -

2.2.12 Ft.micaNG METHOD

The flagging technique involves the use of flannel material or a net designed as in Fig_ 3b. These
materials are flagged from one side of the operator to the other in a semi-circular manner, as one
walks through the vegetation at a slow pace. This should be done in a transect over a specified
distance of about 20-50 m. The activity has to be replicated so as to get unbiased sampling of ticks.
Flagging method is suitable for collecting both adult and immature ticks in higher vegetation.

2.2_3.3 CARBON DroxiDE TRAP

Carbon dioxide is known to attract and activate (especially quiescent) ticks and stimulate questing
postures in them. The best CO, source is dry ice_ This can be put in a perforated container, outside
which a sticky material is applied to trap ticks, CO. method is also suitable for collecting ticks from
thickets and bushes, where dragging and flagging methods cannot be applied.

2_7.3.4 PHEROMONE TRAP

Synthetic attraction-aggregation-attachment pheromone (AAAP) is used to attract conspecific males

and females of Amblyomina ticks to a site. The pheromone is composed of ortho-nitrophenol, methyl
sal cy late and nonanoic acid. It can be placed in a simple container with perforations, outside
which is smeared a sticky material. The effects the pheromone are facilitated by initial
stimulation of ticks by carbon dioxide.

7
 (a)

Wooden base

(b)
Leather material

Handle

Flannel material

3. Methods of collecting host-seeking Licks in pasture: a. dragging method; h. flagging method

8
2.3 Preservation and Labelling of Ticks

When ticks are collected from hosts or vegetation, they should be placed in 70% alcohol in universal
or bijou bottles and the lids (with rubber lining) closed firmly. If possible, ticks should be collected
from different predilection sites, host species and localities and preserved in different bottles. The
engorged females, nymphs and larvae should be collected carefully by hand or using fine curved
forceps to avoid damage. They should be transferred to suitable glass bottles or plastic tubes
individually or in a batch, depending on the species. In case of glass tubes, place moist filter paper
or cotton wool at the bottom of each tube and plug the neck with cotton wool. One end of the plastic
tubes should be cut open and plugged with plaster of Paris which should he kept moist (Kaiser,
1987).

Each bottle of ticks should be labelled with the following information: date of collection, locality
or area, district or province, altitude (whenever possible), host species, age and sex of animal,
predilection site and health condition of host. Labelling should be done in pencil (to avoid being
dissolved in alcohol) on a white glossy paper and placed inside the bottles.

9
3.0 Rearing of Ticks for Identification
It is usually more difficult to identify female ticks than the males because they have limited striking
morphological characters. However, their identification can be made easier if found in association
with males. Similarly, the immature ticks (larvae and nymphs) arc very difficult to identify to
species level, because the development of their structures are not complete. Hence. it is normally
advisable to rear these ticks, under laboratory conditions, to adult stage so as to establish their
identity.

The engorged females, nymphs and larvae that cannot be identified accurately are given tentative
names and maintained in the laboratory at 28DC and 85% relative humidity (RH). Similarly, the
partially fed and unfed ticks that cannot he identified accurately are also given tentative names and
put on rabbit's ears (in cloth bags) to complete feeding.

The engorged females are held individually in tubes, while the larvae and nymphs are held in
batches of about 111-50 depending on the size of the tubes_ They are maintained at 28°C and 85%
RH and observed daily to assess their development processes.

For engorged female ticks, records are kept on the following:

(1) Pre-oviposition period—the lime interval between dropping from the host and laying the first
hatch of eggs

(2) Oviposition period—the time taken for the female to complete egg laying

(3) Pre-eclosion period the time interval between laying of eggs and the first emergence of
—

larvae

(4) Eelosion period•the time interval between the first emergence of larvae and completion of
egg hatching

(5) Hatchability—the proportion of eggs that hatch into larvae. This can be expressed in percentage,
based on the ratio of eggs hatched to total eggs laid

For larvae and nymphs, the following records should be kept:

(I) Pre-moulting period—the time interval between detachment from host to first emergence of
the next developmental stage

(2) Moulting period—the time taken for moulting to be completed.

II)
Some species of ticks are closely related morphologically and may be difficult to distinguish initially
under the dissecting microscope_ One way of confirming these species is to carry out cross-breeding
experiments of males and females, Identical numbers of males of one tick strain and the females of
another strain are applied on the same ear of rabbit and allowed to feed and mate. On the other ear
of rabbit are placed homogenous pairs of ticks of similar strains, to serve as controls. Observations
are made daily on general behaviour, attachment, mating and feeding processes up to engorgement.
Any abnormalities in the feeding processes are noticed,

The engorged I males are weighed and kept individually in tubes at 28°C and 85% RH. The processes
of oviposition and hatching are recorded as explained earlier. If possible, the batches of eggs laid
by each tick may be weighed. The quality and viability of eggs derived from heterogenous crosses
should be compared to those of homogenous strains of ticks.

The larvae derived from cross-bred ticks are reared up to adult stage under laboratory conditions.
The resulting adults are reared to establish the viability or fertility of the offspring of the hybrid.
Whenever interbreeding takes place during the first crossing, there is need to perform a backcrossing
experiment so as to confirm the genetic or cytoplasmic incompatibility. This involves backcrossing
each sex of the hybrid with the parent of the opposite sex, Genetic or cytoplasmic incompatibility
is reflected in the distorted sex ratio (production of males mostly), reduction in the numbers of
offspring or no offspring. Ticks which demonstrate these features are likely to be distinct species.
The data obtained on the productivity (from egg to adult) of hybrid ticks are compared with those
of homogenous breeding, and the appropriate statistical analysis carried out to determine the
significance of differences between them,
4.0 Requirements for Tick Taxonomy
Taxonomic studies, especially the morphological aspects, do not require a lot of sophisticated
facilities. The following are the basic requirements in a laboratory:

(1) Dissecting microscope

(2) Transmission-light microscopy
(3) Glass slide
(4) Wax or putty material for supporting Licks
(5) Forceps
(6) Petri dishes
(7) Filter papers
(8) 70% alcohol
(9) Berlese's fluid (mountant), comprising of:
(a) Water 50 ml
( b ) Chloral hydrate 50 g
(e) Glycerol 20 ml
(d) Gum a r a b i c 30g
(e) Ac e t i c a c i d 5ml
(10) Labels

12
5.0 Introduction to Identification of Ticks

Ticks belong to three famill?s,Argasidae, Ixodidae and Nuttallielidae, which are derived from the
Following taxonomic tree:

Phylum: Arthropoda
Class: Arachnida
Order: Acarina
Sub-order: lxodoiden
Argasidae
lxodidae
Nuttallielidae

Cicncra: Argas, Rhipicephalus, Amblyonuna, etc.

Species: Argus persicus., Rhipicepholus appendiculaws, etc.

5.1 Differentiation of Major Groups of Ticks

The first step in the identification of ticks is to distinguish between the major groups: families,
genera, sexes and developmental stages, before attempting to identify to species level. This can be
achieved by following the simple procedures modified from Kaiser (1987). The main distinguishing
features between these groups of ticks are provided later for ease of reference.

The procedure for identifying ticks is as follows:

(I) The preserved specimens of ticks collected from the field are poured into a Petri dish and
separated from foreign material such as hair, scales or dry skin. They are spread on filter
paper, then blotted to absorb excess preservation fluid.

(2) Ticks which are dirty should have their scutums cleaned by gently rubbing them on filter
paper.

(3) Each tick is placed on clean filter paper in a Petri dish on the stage of a stereoscopic dissecting
microscope and exposed to adequate illumination. Specimens should not be allowed to dry
completely, as their legs may break. and occasional dampening with 70% alcohol may be
required.

(4) Use the low power objective to separate genera and sexes of ticks. Use the medium or high
power objectives to identify to species level.

13
 (5) Use Forceps to manipulate ticks and tilt them towards the light source or put them on one side.
so as to clearly see the key diagnostic features. Use plasticine or a ball-borne stage to support
the ticks.

(6) When identifying the female ticks. the characteristics of the female genitalia can sometimes
give a conlirmatory identification. Refer to Annex 1. describing a rapid method for cleating
and 11101.11ai nsi, of the genital apron of the ixodid tick (Kaiser, 1987).

5,1.1 Differences Between Argasidae *Nand Ixodidoe (Haul) Ticks

The morphological differences between soft and hard ticks can be seen by comparing Figs 4-7.
Figures 4 and .5 show the morphology of two soft tick genera. while Figs 6 and 7 illustrate the key
identification Features of hard ticks. The basic distinguishing features are described in Table I.

5.1.2 Genera of Ticks

The family lxodidue has 13 genera and 671 for more) species of ticks, the Argasidae has 5
genera and about 170 species of ticks_ while the family Nuttallielithie has only one species. The
number of tick species in each geucra is provided for the family Ixodidne as shown in Table 2,

The Ltenera of ticks in the family Isodidae can be differentiated easily by using the Flow Chart I (p.
21). In addition, the key identification Features for each genus of ticks arc highlighted in Fig. 7 and
Plates 1 1 6. (Note that Plates 4 1 6 Lire found in the centre section or colour plates'.
—

I-1
Table 1. Basic morphological features used for distinguishing between soft and hard ticks
Morphological
characters Argasidae Ixodidae

Not sclerotised Sclerotked whole dorsal

Scutum aspect of male. but about a
third of clorsum of female

Texture smooth and glossy Dark brown/brown and in

Cuticle some cases ornamented

Located away from anterior end of Located at anterior end of

Capitulum tick, not visible dorsally body and visible dorsally

Less developed—does not facilitate Strongly developed with

Hypostome firm and long attachment firm retrograde teeth
(chelicera)

Absent or present in lateral Absent or present and

Eyes exposed near lateral
folds
margins of scutum

Small, located in unique Large, located posterior

Spiracular plates positions anterior to coxae IV to coxae IV

Absent Present or absciit

Festoons
Absent Present and located in
Ventral plates various positions

Located more or less between Located between first and

Genital aperture first pair of coxae second pairs of coxue

'5
 Table 2. The genera and number of tick species in the family
Ixodidae
Family Genera Number of species
Ixodidae A/nblyomma 102
Anomalohimalaya 3
A p ► ► o ma 24
Boop► ilus 5

Dermacentor 30
Hatunaphysalis 155
Hyalomma 30
Lodes 245
Margaropus 3
Nosomma 1
Rhipice►tor 2
Rhipicepliaius 70

Total 671

16
 DORSAL VIEW
Fig. 4. Genus Argas: Key morphological characters used in identification. (Adapted from floogstraal,

VENTRAL VIEW

Capitulum
(mouthpart)

Genital aperture

Differentiated periphery
(lateral suture)

1956)

17