AFRICAN WILD DOG (LYCAON PICTUS)

BREEDING PROGRAM Mkomazi Game Reserve, Tanzania

Report 2001-2005

Aart Visee, D.V.M.

AN ENDANGERED SPECIES PROGRAM UNDERTAKEN BY:

THE WILDLIFE PRESERVATION TRUST FUND (TANZANIA)

THE GEORGE ADAMSON WILDLIFE PRESERVATION TRUSTS

(UK,USA, NETH., GER., KEN.) and THE AFRICAN WILD DOG

FOUNDATION (NETH.)

2

CONTENTS Page

Foreword 3

Introduction 4

Husbandry 5

Pack composition, development and behaviour 5

Introduction 5

2001: Restart / Malaika pack 5

2002: Kimondo pack / Start Zawadi pack 7

2003 9

2004 10

2005 14

Zootechnique 18

Identification 18

DNA 20

Housing 21

Nutrition 21

Bodyweight and Neck size 22

Veterinary 25

Preventative Medicine 26

Canine Distemper Virus 26

Rabies 29

Parvovirus 31

Parasites 31

Clinical Work-Diseases 33

Clinical Work 33

Anaesthesia 34

Pathology 35

Acknowledgements 36

Summery 37

References 38

Appendix I, Bodyweight and neck size 40

Appendix II, Distemper VNT 41

Appendix III, Distemper Elisa 42

Appendix IV, Rabies 43

Appendix V, Pack composition December 2005 44

Appendix VI, Breeding results 2001-2005 45

Appendix VII, Breeding results 2001-2005 including similarity 46

Genetic Analysis 47

3

Foreword.

With lots of enthusiasm and high hopes, we started the African wild dog breeding program in

Kisima camp, Mkomazi Game Reserve, Tanzania, in 1995. The future looked bright with 25

beautiful pups, and we were determined to do something to preserve the African wild dog

from extinction by way of breeding them and reintroducing them to the wild, in places where

they had disappeared or seemingly disappeared.

The first 5 years of our breeding program were a success. We expected some difficulties, got

them, and were able to solve them. But, we never expected to be confronted with a very

serious canine distemper virus (CDV) outbreak, that hit our African Wild Dog Program, by

the end of 2000. At that time the program contained 52 dogs. By the time the virus had had its

way only three dogs were left. A really devastating experience, not only for the dogs but for

the people involved as well.

In fact, this outbreak symbolized the end of the first five years of our breeding program. This

period ended abruptly, and completely when the last of the 25 original dogs died August

2001. At about the same time, a new era began on the seventh of July 2001, symbolized by

the birth of the first litter of pups to the dogs, which had survived the canine distemper

outbreak.

The years 1995 to 2001, including the canine distemper epidemic, were extensively reported

in the Report 1995-2001. Since then no new report appeared, as there was not much to report,

being quite and thus nice years for a change. The establishing of new breeding packs with

subsequently the birth of pups, and the progress made in the canine distemper vaccination

research made it worthwhile to come out with a new report covering 2001-2005.

Aart Visee

Aart M. Visee. D.V.M.

The African Wild Dog Foundation

Gaspeldoornlaan 17, 3956DN Leersum, The Netherlands

Tel.: +31 343 769081

E-mail: aartvisee@gmail.com

4

Introduction.

The George Adamson Wildlife Preservation Trusts, (joined in 2000 by the African Wild Dog

Foundation), through their Field Director Tony Fitzjohn, were invited in July of 1988 by the

Tanzanian Government, to establish a rehabilitation program for the African wild dog

(Lycaon pictus) involving capture, captive breeding, and reintroduction.

The African wild dog is an extremely endangered species. Diseases, large predators (lions),

and man threaten their existence. In areas where game is scarce, wild dogs tend to get close to

human settlements. As a consequence they are poisoned, and may come into close contact

with domestic dogs. As the African wild dog is susceptible to diseases also found in domestic

dogs (distemper, rabies and parvovirus), cross infection may occur; leading, in some cases, to

high mortality.

It took many years, and much searching before dens were finally located from which pups

could be collected. It was most important that the pups came from a non-conservation area (in

this case the Masai Steppe), to prevent the unnecessary decline of numbers in conservation

areas. And, it was thought likely that pastoralists would have poisoned them sooner or later

anyway. Finally, the right moment arrived in early August of 1995. Twenty-five pups were

lifted from three different dens in the Masai Steppe. At the time of lifting, they were not

totally dependent on their mothers--they could eat solid food. They ranged in age,

approximately, from three to five weeks.

For them, and the people involved, it represented a new start for the rescue of the genetically

unique East African population of the African wild dog (Lycaon pictus)--considered by some

researchers to be a distinct subspecies. With the arrival of the dogs, it was decided to collect

as much data as possible without stressing the dogs too much, and with a minimum of

interference in their lives. For these reasons, blood samples were not taken on all occasions

from each of the dogs. Alpha females, and alpha males were sedated as little as possible, to

prevent any possible adverse effects on their social status. Pregnant females were not sedated

for obvious reasons.

For 5 years the program developed well, and prospered. By the end of 2000, there were a total

of 52 dogs, and plans for re-introduction into the wild were seriously underway. However,

just before Christmas 2000 fate struck. The breeding program was hit very hard with a canine

distemper virus infection (CDV), despite the fact that the dogs had been vaccinated according

to an intensive schedule. The Mkomazi African wild dog population was almost completely

wiped out, and in 2001 it was needed to start all over again with the three survivors as a basis

for a new population. Soon two “volunteers” dogs would join them. Together with one of the

distemper survivors, and the first litter born out of the distemper survivors, they would form

the nucleus of our new breeding program.

This report is divided into two parts, Husbandry and Veterinary.

5

HUSBANDRY

Pack composition, development and behaviour. This report includes how the new breeding packs were composed and developed. Where

necessary behaviour is reported and commented on.

Zoo technique.

In this section, a report is drawn up about identification, DNA, housing, nutrition, bodyweight

and neck size.

Pack composition, development and behaviour (see appendix VI)

Introduction

The key to understanding how new free living wild dog packs form and persist, despite major

changes in their composition in subsequent years, is the existence of a basic protocol in wild

dog behaviour, that was first identified in the East African wild dog population to which the

Mkomazi captive breeding group belong.

In both the breeding and feeding hierarchies older dogs of both sexes (apart from the alpha

[i.e. main breeding female - the ‘Selfish Matriarch’ (Burrows 1992)] give priority to both

related and unrelated younger dogs in the pack (Malcolm & Marten 1982, Burrows 1995 &

2004, McNutt, 1996, Visee et al 2001).

In this report the term ‘natural rules’ is applied to any behaviour observed in the Mkomazi

captive packs that is known to be typical of the behaviour of free living dogs.

2001: Restart / Malaika Pack

After the devastating canine

distemper period from December

2000 – February 2001, it was a real

thrill for all the people, and dogs

involved, to see the three survivors

united on March 13th 2001. During

the preceding weeks they had been

calling to each other every night.

For the occasion the two males,

#325 (born 4-9-97) and #372 (born

8-1-99), were sedated and

transferred to the feeding area of

the Sangito boma. The female

#306 was locked out of the feeding area. The moment she noticed the other dogs, still sedated,

she started to whine softly. After waking up, the two males (seeing each other for the first

6

time) showed no signs of hostility toward each other, although #325 took more time for a

complete recovery.

In the meantime, #306 tried to make contact through the wire, and started to roll over onto her

back. After 2½ hours, #325 was completely recovered, and since no signs of hostilities were

shown through the wire (in fact #306 and #372 were very interested in each other), the dogs

were put together and the Malaika pack was established.

Immediately female #306 and the younger male #372 joined, and #306 was putting her nose

under #372’s belly. #325 kept a little distance. It appeared that the female chose the younger

male.

The next day, the three of them were

eating and sleeping together.

Apparently at the introduction, it was

already decided who was going to be

alpha male, #372, being the youngest

of the two males. In the second half of

April, #306 and #372 mated, and on the

7th of July, #306 gave birth to eight

pups (m5, f3). The pups flourished.

On the 4th of August, fate struck again

when suddenly female #306 showed

signs of serious distress and died within

six hours (see Report 1995-2001). That

day, the pups were exactly four weeks of age, and old enough to live on solid food. Having a

lot of experience in raising the original pups, it was decided to separate the pups from the

males, to keep a close watch on them. The pups were transferred to the Kisima boma.

Nevertheless, one female pup died on the 3rd of October (see Report 1995-2001). The

remaining seven pups grew up well.

The Malaika pack was short lived but left 7 beautiful pups as a strong basis for the breeding

program.

At the end of June, two female wild dogs showed up in the vicinity of our boma’s. It was

most likely that they were two young sisters (18-24 months old) who had emigrated from

their free living natal pack and were looking for an all-brother party to start a new pack. The

females were very interested in the pups as well; they kept coming and going for the next

couple of months.

After the death of #306 they stayed even more closely to our boma’s. Apparently, they had no

intention of leaving, and the decision was made to try and introduce them to our males. At the

end of August the door of one boma was kept open and the two females went in. Before the

door could be properly shut one female escaped. However, she stayed close to the boma

where her sister was kept. Toward the end of September she was still hanging around, and it

was apparent that she had no intention of leaving. At that time the wildlife veterinarians from

TAWIRI (Drs. Robert Fyumagwa and Harald Wiik) were in Mkomazi on other duties, and

they decided to provide us with their professional assistance in order to transfer her safely into

7

the boma; finally reuniting her with her sister. Few hostilities took place when the four dogs

were introduced to each other.

In December 2001, it was apparent that #372 was the alpha male, and #040 the alpha female.

Normally in a free living pack which contained younger males it might have been expected

that after the death of the alpha female (#306) the alpha male would also have lost his

position. As in this case there was no younger male, only his older male pack member, he

kept his alpha position. If there had been a younger male, #372 might have lost his alpha

position (based on the ‘young male first’ protocol in wild dog society) when new packs are

formed.

Pack composition December 2001:

KISIMA boma

Females Males

040 alpha, Kimondo 372 alpha, Chupu chupu

006, Zawadi 325, Bahati

SANGITO boma

pups born 07-07-01 (306 x 372)

Females Males

448 202

464 251

269

368

389

2002: Kimondo pack / Start Zawadi pack

January 2002 mating was observed between #040 (Kimondo) and #372 (Chupu chupu),

giving evidence that they were indeed the alpha’s. #325 (Bahati), the other distemper male

survivor, was subordinate to #372 from

the moment these two were put

together in the same boma in March

2001. While #325 was fast asleep he

was bitten in the neck many times by

#372. The fact that they originated

from different packs might have

contributed to this kind of behaviour.

March 20th #325 the subordinate male

died suddenly 5 minutes after eating

from, what appeared to be, heart

failure.

8

Kimondo (#040) gave birth 18 April. Some weeks later it appeared to be a litter of 4 pups.

Four days later(22 April) Zawadi (#006) gave birth to four pups as inspection of the den

showed a couple of days later. It was observed that both females had mated with Chupu

chupu (#372).

In the following days Kimondo (#040) went in

Zawadi’s den on a number of different

occasions. Inspection of the den on 29 April

revealed no signs of pups. Thereafter Zawadi

assisted in raising and feeding Kimondo’s pups,

being an integral part of the upbringing of the

pups as it should be. When the pups were about

6 months old, Kimondo and Chupu chupu

started to chase Zawadi off and as a result she

kept herself more and more on the side.

Kimondo rarely bit her sister but just chased her

off until she stayed away.

In free living packs, subordinate sisters of the alpha female often leave (re-emigrate from)

their alpha sister’s pack, having assisted in raising a new packs first litter, and attempt to

locate new males to form a new pack.

Following these free living pack procedures, Zawadi was removed on December 8th and

moved to the Lendanai boma to start a new pack with the brothers #202 and #251 from the

Malaika litter in the Sangito boma. In this way a new pack, Zawadi pack, was established. To

prevent brother sister relationships the other 3 males from the Sangito boma, #269, #368 and

#389 were moved to the Ayubu boma.

Pack composition December 2002:

KISIMA boma

Kimondo pack Females Males

040 alpha, Kimondo 372 alpha, Chupu chupu

Pups born 18-04-02 104 651

348 730

LENDANAI boma

Zawadi pack Females Males

006, Zawadi 202

251

SANGITO boma

Females

448

464

9

AYUBU boma

Males

269

368

389

2003

Kimondo pack: May 31st Kimondo (#040) gave birth to six pups; four males and two

females. There was a lot of excitement from the first litter of pups (now 14 months old

yearlings) and they all assisted in feeding and raising the new pups. At the time the new pups

started to eat non-regurgitated food in the feeding area, the yearlings hung back and the alpha

male (#372), chased them back as well, just to make sure.

An experiment in 1998 in Mkomazi, to test whether the ‘natural rules’ observed in the wild

applied equally in captivity, showed that the order of feeding and the time spent feeding at the

food provided (i.e. as at the kill of a free living pack) were very similar. In both cases pups

monopolize food often with the alpha female (the ‘Selfish Matriarch’) and defended the

food/kill from the other adults and only after the pups are satiated do the older dogs return to

the food with older subordinate dogs the last to feed.

The time spent by the Mkomazi captive pack feeding on the food provided was 40 minutes at

one feed and 50 minutes at another with the pups (4) monopolizing the food for 35 of the 50

minute feeding session. This is similar to the time taken by free living Serengeti packs with

pups and/or yearlings spent feeding (‘Access Time’) at a kill (Schaller 1972, Malcolm 1979,

Fanshawe et al 1993). There is no ‘scramble competition’ for food (contra Carbone et al

1997) and dogs of different ages/cohorts do not feed equally at any one feed/kill. This ‘natural

rule’ (the ‘feeding’ or ‘access’ hierarchy’ protocol) is now known to apply to both captive

and free living packs. This is important as we know that the ‘natural rules’ will be followed by

Mkomazi bred dogs post release.

When the new pups were approximately six months old the two

male yearlings started to snap at their father, #372, at feeding

time. They attacked him together from each side. As a result

#372 made sure that he quickly was in the feeding area first, had

a quick bite and hung back to allow the pups and the yearlings to

eat. Kimondo did not have any real problems with any of her

pups or yearlings. She drove them away if they attempted to stop

her feeding with them.

Zawadi pack: The initial introduction of the dogs to each other

was peaceful, although the dogs were periodically somewhat

exited. They settled down in their own way, i.e. Zawadi (#006),

was much shyer and as a consequence more nervous and

remained most of the time on the periphery. Until October both

males, #202 and #251, chased and snapped at Zawadi

occasionally, although #202 tried to get closer to her and sometimes even played with her.

10

Most of the times, however, #251 interfered and chased Zawadi away. At this time it was

difficult to tell who was the alpha male. Originally, in the litter, #202 appeared to be alpha. As

a consequence Zawadi had to be feed separately to make sure that she got enough food. On

the rare occasions that they played together Zawadi was always at the back, nervous and

never really involved.

By October #251 started to come closer to Zawadi, however #202 started to chase her off

when she came too close to #251.

Sangito boma: The only two dogs in this compound, the two sisters, #448 and # 464, had an

uneventful year. In 2002 it looked like #464 was alpha, but in the course of 2003 it showed

that #448 was alpha. At feeding time she chased #464 off but outside feeding time they were

very close.

Ayubu boma: Over the year the positions of the three

brothers in this boma were very clear; #368 is the alpha,

being very fierce and displays it to his brothers. #269 is

the most subordinate in the pack. He did not display

much physical strength but made the most noise (typical

for submission): he was the one who started to “bark”

when people or some animal approached the boma. As a

result of their clear dominant/subordinate relationship

they did not fight each other.

Pack composition December 2003:

KISIMA boma

Kimondo pack Females Males

040 alpha, Kimondo 372 alpha, Chupu chupu

Pups born 18-04-02 104 651

348 730

Pups born 31-05-03 001 092

031 187

212

393

Lendanai, Sangito and Ayubu boma as December 2002

2004

Kimondo pack: In June Kimondo, #040, again came in season and on August 29th gave birth

to her third litter that appeared to contain six pups. When they were old enough to come out

of the den it appeared as if they were going to be fed by quite a number of dogs.

However, already during pregnancy Kimondo did not look well and after coming out of the

den the pups remained very tiny and did not increase in weight or size. The pups of Kimondo

II litter (now 14 months old yearlings) hassled them. Although the yearlings fed them they

11

also picked them up by the ears or the middle of the back, not by the neck and were throwing

them around. Also, while trotting after their parents along the fence, they were trampled upon

by the yearlings. The first three pups died 11 October and the last three died on October 23rd,

i.e. they disappeared and no remnants were found.

The intriguing facts here are that the dogs of the first born litter, Kimondo I (now 28 months

old), ignored the small pups and the dogs of the second born litter, Kimondo II (now 16

months old), hassled them in a rougher way than we observed in other equal events. If there

was going to be any trouble it would have been expected from the dogs of Kimondo I litter.

The removal of these four dogs of the first litter was delayed as it was not advisable at that

time to start a third and fourth breeding pack for which the dogs were intended. The yearlings

of Kimondo II litter were underweight and did not have the body size expected for their age.

It is possible therefore that the apparently

aggressive behaviour of the yearlings toward

the new pups was due to lack of food but this

is unlikely as they did feed the pups. Also as

their mother appeared unwell during her

pregnancy and the pups failed to grow as

expected perhaps the pups were ill.

Yearlings are fascinated by new pups and

attempt to interact with them as much as

possible, so there are many accounts of free

living yearlings appearing to hassle new pups.

If the pups are healthy and well fed they just

retreat into the den and survive despite the

rough play of the yearlings.

There is just a single case reported in a free

living pack when yearlings reportedly did not

feed young pups at a den and even stole food

regurgitated to the pups by other dogs. This

was a pack watched in the dry season in

Serengeti when prey was scarce and the pups

were obviously both starving and in very poor

health (Frame & Frame 1981). This was a time in Serengeti when canine distemper was

suspected as the cause of high mortality in pups and perhaps in some young adults (Frame et

al 1979). It may be that these free living pups were not behaving ‘normally’ (as perhaps were

the pups in the Kimondo pack) and had no appetite when fed regurgitated food by the adults,

so the yearlings took any uneaten food.

Not removing these four dogs meant that natural pack rules were not followed and might have

caused trouble, which did not happen. That leaves the abnormal behaviour of the dogs,

yearlings, of the Kimondo II litter. Looking at the dogs of the Kimondo II litter, underweight

and not a body-size according to their age, it still might have been possible that their

behaviour towards the newborn pups was instigated by lack of food. Not being full-grown yet

and receiving not enough food for their own growth the newborn pups were a direct threat to

their own existence.

12

Early December the four dogs of the Kimondo I litter were moved from the Kimondo pack to

the Sangito and Ayubu boma.

Zawadi pack: Between late December 2003 to January 2004 #251 stayed close to Zawadi.

#202 still chased her at feeding time but normally they all slept near each other and were

getting along well in a more normal way.

However, due to her nervousness, if one of

the males woke up, Zawadi immediately

followed him. Early in February it was

observed that the three dogs were running

together more often and by the end of the

month Zawadi came in season. She started

to pursue #251 and #251 also pursued her.

Both chased off #202, with the male #251

the most vigilant and active chaser of

#202. The alpha pair, #006 and #251

showed all the behaviour one might expect

from an alpha pair, i.e. joint urine marking with raised leg posture, dominance display with

#202 performing appeasement rolling, cringing and whining with ears back and tail between

legs.

On May 11th Zawadi gave birth to nine pups, very unusually all were males.

Towards the end of the year the beta male, #202, was chased away at feeding time. However

#202 had enough confidence in the keeper to come in and have some food before the other

dogs were allowed to come into the feeding area.

Sangito boma: On the third of December the males, #651 and #730, from Kimondo’s

firstborn litter were moved from the Kisima boma to the Sangito boma. The dogs were

sedated and woke up in the feeding area of the Sangito boma. The resident females were

locked out of the feeding area. Immediately the dogs showed an interest in each other but with

some reservation in their behaviour. In the morning of 5 December, the day after the transfer

of the male dogs, the dogs were allowed to join each other. No hostilities whatsoever took

place and at feeding time all the dogs fed together.

From the beginning, male #651 and female #464 took up alpha position. The relationship

between the males and the females was pretty relaxed from the beginning. The alpha pair

started mating within a couple of weeks.

Ayubu boma: On the third of December the females, #104 and #348, from Kimondo’s

firstborn litter were moved from the Kisima boma

to the Ayubu boma. The dogs were sedated and

woke up after transport in the Ayubu boma outside

the feeding area. The resident males were locked up

in the feeding area. In this way the females had the

opportunity to inspect and get familiar with their

new housing arrangements should the males be

aggressively interested in them. The opposite

happened. The females showed more interest in the

males than their brothers did in the females in the

Sangito boma. The females slept and rested against

13

the fence of the feeding area to be as close as possible to the males. In the morning of 5

December the dogs were allowed to join. Here too no hostilities took place and the dogs

immediately started to act as a pack.

The male #368 behaved as the alpha male from the start and took great interest in the

females, following them around. The female #104 appeared to be the alpha female from the

start. The other two males had to keep a subordinate position which meant that they had to eat

in a secondary pattern to the alpha pair. The alpha pair has been seen mating within a couple

of weeks.

Pack composition December 2004:

KISIMA boma

Kimondo pack Females Males

040 alpha, Kimondo 372 alpha, Chupu chupu

Pups born 31-05-03 001 092

031 187

212

393

LENDANAI boma

Zawadi pack Females Males

006 alpha, Zawadi 202

251 alpha

Pups born 11-05-04 666

671

732

853

860

887

926

939

949

SANGITO boma

Females Males

448 alpha 651

464 730 alpha

AYUBU boma

Females Males

104 alpha, Mrefu 269

348 beta, Regina 368 alpha

89

14

2005

Kimondo pack: After the removal of the members of the Kimondo I litter the stress during

feeding time completely died down and mid January it was observed that the alpha pair was

mating again. During pregnancy Kimondo was very vocal and at feeding time she whined

endlessly to ensure that the other dogs regurgitated their food for her. She gave birth 10 th

April to six pups, two males and four females. From the moment the pups emerged from the

den a female pup appeared to be smaller and weaker than her littermates. Lacking in physical

strength at feeding time she had to struggle to get some food and as a consequence was

steadily falling back in strength and size in comparison to her littermates. July 17th she gave in

and died. The other five pups grew up well and behaved in a normal way, thus taking and

being allowed first place at feeding time.

On 26th November the four males of the Kimondo II litter were removed from the pack and

transferred to the Kilo Sierra compound in the rhino sanctuary, to be the first dogs to be

released.

Zawadi pack: Mid February Zawadi #006 and #251, the alpha pair, was mating again.

Zawadi too was very vocal during pregnancy and also tried to encourage the other dogs to

regurgitate food for her. The

subordinate male #202 was the only

one to do so. She gave birth 22nd May

to five pups; once again all males.

After the pups emerged from the den

the nine pups from the older litter

assisted in feeding them as it should

be.

On 25th November the nine male

pups, Zawadi I litter, were transferred

to a newly build boma as they had

reached an age that usually free living

yearlings leave their natal pack. They

settled well with hardly any calling at night.

Sangito boma: Despite the observed mating in December 2004 and the display of aggressive

behaviour by the alpha male #730 towards his brother and the alpha female chasing off her

sister it appeared that the alpha female #448 was not pregnant.

The feeding system had to be manipulated because the alpha pair would otherwise leave the

beta dogs with very little food.

The following interactions were observed: At the time it was presumed that the alpha female

was pregnant, she chased her sister out of the group. However, as the months passed, the beta

sister returned slowly to the group, but still kept some distance. When the dogs were trotting

through the compound, the alpha pair was first in line, followed by the beta male, and slightly

behind the beta female. Also, when they slept, the alpha pair and the beta male slept together,

and the beta female slept apart.

15

Ayubu boma: Since the alpha female #104 became pregnant December 2004 the other two

subordinate males in the compound (#269, #389) had been experiencing problems. The alpha

male continually chased them off during eating and they had to be fed separately. However,

the subordinate female #348 was chased off by the alpha female for the first couple of weeks

of pregnancy, followed by a couple of weeks that she was accepted again and the last couple

of weeks of her pregnancy #348 was chased off again.

March 27th #104 gave birth to 7 pups. As in Kimondo’s litter one pup was half the size of the

others at the first sighting and died 31st May.

Of the six left, three were male and three were female. Another female pup appeared to be

retarded in growth and died July 17th.

Mid June mating was observed by the alpha male #368 and beta female #348. From that

moment on the alpha female #104 chased her subordinate sister off.

The subordinate beta female #348 gave birth on 6th August 2005. The pups were for the first

time sighted on 19th September and

appeared to be seven pups in all.

When #348 was pregnant, the alpha

female chased her off at feeding time

and tried to keep the alpha male

away from her. But since she gave

birth, the alpha female allowed her

to feed normally, i.e. allowed her to

come to the food in the feeding area.

Then #348 went around all the adult

males whining and succeeded in

getting them to regurgitate for her.

She ate that food as well and then

regurgitated for her pups.

Meanwhile the alpha female, #104, looked after her own pups without getting any assistance

from the adult males in the compound. The alpha male too assisted in feeding the pups of the

beta female.

The pups from the alpha female were very curious to see what was inside the den of the beta

female, as they could hear the noises and cries. They often went down into the den and came

out the other side, as the new pups (beta) were hidden in a channel off the main den.

The alpha male was fierce in protecting both litters of pups but spent most of his time with the

beta female #348.

On 26th November a male pup of the litter of the beta female died.

16

Pack composition December 2005 (see also appendix V):

KISIMA boma

Kimondo pack Females Males

040 alpha, Kimondo 372 alpha, Chupu chupu

Pups born 31-05-03 001

031

Pups born 10-04-05 039 043

2040 073

046

LENDANAI boma

Zawadi pack Females Males

006 alpha, Zawadi 202

251 alpha

Pups born 11-05-04 666

671

732

853

860

887

926

939

949

Pups born 22-05-05 519

521

560

567

572

SANGITO boma

Females Males

448 alpha 651

464 730 alpha

17

AYUBU boma

Females Males

104 alpha, Mrefu 269

348 beta, Regina 368 alpha

389

Pups born 27-03-05 444 557

481 586

643

Pups born 06-08-05 000 232

6372 296

0348

2368

18

Zootechnique

Identification All the dogs when first sedated were fitted with a transponder. The transponders were inserted

subcutaneously on the left side of the neck. The transponders were checked on every occasion

the individuals were sedated. All appeared to be in good order. The dogs received

transponders starting with the same number 528210000 for every dog, followed by a different

six number identity for each dog. Figure 1 shows the code for each dog. The only exception is

#372, the last living survivor of the canine distemper epidemic. His complete number is:

981093500110372.

To make the numbers more workable only the last three numbers are used on daily basis.

As an extra precaution photographs were taken from both sides of the dogs in lateral

recumbency.

19

Table 1: Transponders Number

ID number

Litter of origin

Sex

Date of Birth

Date transponder Applied

181006 006 F 1999 20-11-01

181040 040 F 1999 20-11-01

180202 202 Malaika M 07-07-01 22-11-01

180251 251 Malaika M 07-07-01 22-11-01

180269 269 Malaika M 07-07-01 22-11-01

181368 368 Malaika M 07-07-01 22-11-01

180389 389 Malaika M 07-07-01 22-11-01

180448 448 Malaika F 07-07-01 22-11-01

180464 464 Malaika F 07-07-01 22-11-01

181104 104 Kimondo I F 18-04-02 05-12-02

180348 348 Kimondo I F 18-04-02 05-12-02

179651 651 Kimondo I M 18-04-02 05-12-02

179730 730 Kimondo I M 18-04-02 05-12-02

268001 001 Kimondo II F 31-05-03 04-12-03

269031 031 Kimondo II F 31-05-03 04-12-03

273092 092 Kimondo II M 31-05-03 04-12-03

269187 187 Kimondo II M 31-05-03 04-12-03

268212 212 Kimondo II M 31-05-03 04-12-03

266393 393 Kimondo II M 31-05-03 04-12-03

269666 666 Zawadi I M 11-05-04 04-12-04

266671 671 Zawadi I M 11-05-04 04-12-04

265732 732 Zawadi I M 11-05-04 04-12-04

266853 853 Zawadi I M 11-05-04 04-12-04

266860 860 Zawadi I M 11-05-04 04-12-04

268887 887 Zawadi I M 11-05-04 04-12-04

267926 926 Zawadi I M 11-05-04 04-12-04

264939 939 Zawadi I M 11-05-04 04-12-04

272949 949 Zawadi I M 11-05-04 04-12-04

268444 444 Mrefu I F 27-03-05 24-11-05

268481 481 Mrefu I F 27-03-05 24-11-05

272557 557 Mrefu I M 27-03-05 24-11-05

269586 586 Mrefu I M 27-03-05 24-11-05

269643 643 Mrefu I M 27-03-05 24-11-05

788039 039 Kimondo IV F 10-04-05 26-11-05

802040 2040 Kimondo IV F 10-04-05 26-11-05

788043 046 Kimondo IV F 10-04-05 26-11-05

794046 043 Kimondo IV M 10-04-05 26-11-05

805073 073 Kimondo IV M 10-04-05 26-11-05

789519 519 Zawadi II M 22-05-05 25-11-05

796521 521 Zawadi II M 22-05-05 25-11-05

795560 560 Zawadi II M 22-05-05 25-11-05

786567 567 Zawadi II M 22-05-05 25-11-05

792572 572 Zawadi II M 22-05-05 25-11-05

Number all transponders start with 528210000 followed by the number in the first column

20

DNA (see also appendix VII)

November 2002 blood was collected and DNA isolated from the following dogs:

Malaika litter, born 07-07-01, parents 306 (F) and

372 (M):

464, 448, 251, 368, 269, 202, 389

Wild caught: 006 and 040

Bred in Mkomazi:

372, parents 284 (F) and 298 (M)

325, parents 262 (F) and 274 (M), died March

2002, no off-spring

December 2004 blood was collected and DNA isolated from the following dogs:

Kimondo I litter, born 18-4-02, parents 040 (F) and 372 (M):

104, 348, 651, 730

Zawadi I litter, born 11-05-04, parents 006 (F) and 251 (M):

666, 671, 732, 853, 860, 887, 926, 939, 949

The DNA was isolated by Gendika BV, Veendam, the Netherlands.

Gendika BV performed DNA fingerprinting on the DNA of the above-mentioned dogs (see

attached report). From the results could be concluded that there is still a considerable

genetic variation in the African wild dog population in Mkomazi. Answers were given to the

following questions:

a. It was assumed that the new dogs #006 and #040 as they arrived together were sisters.

The similarity between these two dogs proved to be 0.78, which is quite high and

makes it most likely that they are sisters indeed.

b. What is the genetic relationship between the new dogs, #006 and #040, and the

original dogs (v.d.Zande1997)? Apparently the relationship is not high and these

females could have bred very well with the original dogs especially dogs from the

Lendanai group. Unfortunately all the original dogs, except three, died in the

distemper outbreak. Of these three dogs only male #325 descended from the Lendanai

but died one year after the outbreak bringing the Lendanai line to an end.

c. What is the genetic relationship between the new dogs, #006 and #040, and the last

living off-spring, #372, of the original dogs (being father of the Malaika litter and

Kimondo I litter)? The similarity proved to be rather high, higher than the similarity

between the three groups of original dogs in 1995.

d. Is it acceptable to combine pups born out of the combination #372 x #306 with pups

born out of the combination #372 x #040? It appeared to be that all combinations of

different nests is possible as all combinations show lower similarity with other

combinations than the similarity calculated from within nests.

e. Would it be genetically acceptable for re-introduction purposes to mix dogs born out

of the combination mentioned under d with pups born out of the combination #006

21

and #251? This question is already answered under d, although there is a preference to

combine pups out of combination #006 x #251 with pups out of the combination #040

x #372.

f. Are there two sub species of African wild dogs as researchers originally claimed

(Girman et al 1993) but later on further investigation changed their minds (Girman et

al 1997)? The similarity between the two groups (Mkomazi dogs and Amsterdam Z00-

Artis- dogs) proved to be 0.26 and the conclusion might be justified that there is a

different genetic basis in the Artis wild dogs (Southern Africa originated) compared to

the Mkomazi wild dogs (Eastern Africa originated) but not sufficient to conclude that

there are two subspecies.

Housing

The original four boma’s of the dogs were

not altered. (For details see Report 1997-

2001). The Lendanai boma was

completely stripped for considerable

maintenance in 2002.

As an extra precaution to keep small

animals and people at a distance, a six foot

high fence made of chicken wire was

placed at a distance of approximately two

meters from the outside of the perimeter

fence of the Sangito and Kisima boma’s in

2001, the Lendanai boma in 2002 and the Ayubu boma in 2003.

Two new boma’s were built by the end of 2005 to provide accommodation for the yearlings

that had to be separated from their natal pack due to following natural pack rules. The new

boma’s were constructed in the same way as the other boma’s and have the same size, 70 x 40

meters.

Nutrition

The dogs were fed once a day. As a basis they

received a commercial dog pellet, which was

softened through soaking in water. Mixed

with this dog food were pieces of cow meat.

Once every fortnight two cows were bought

at the local market and slaughtered, that is to

say, as much meat as possible was taken from

the cows and put into a freezer, operating on

paraffin, as much as the freezer could contain.

The rest of the carcass was fed to the dogs.

On the day they received the carcass, they did

not receive the pellets.

22

Due to the fact that the dogs were rather lean December 2004 the temporary measure was

taken to buy two cows every week and thus the quantity meat they received was increased. In

November 2005 the pups born in 2005 showed once again a disappointing bodyweight as well

as the dogs of the Zawadi I litter, born in 2004. As the number of dogs had grown in the

course of 2005, the quantity of cows was increased to three cows a week.

Pellets used were initially GILPA’s “Valu

Complete”, a complete adult dog food. The

moment the pups started to eat solid food, they

received the same softened pellets with cow

meat. The pups grew with no obvious bone

abnormalities.

November 2002 the commercial dog food pellet

brand had to be changed due to the

impossibility of getting an export license in the

UK for Tanzania. After an intensive search

“Bobtail Classic” [Foodcorp (Pty) Ltd, SA] was selected, having the best quality price ratio,

to replace “Valu Complete”.

The dogs were more eager to eat the new pellets than the old ones.

Bodyweight and Neck size (See appendix I)

It was our policy to take the bodyweight and the

neck size of a dog every time the dog was sedated

for whatever reason. The table below gives the

average bodyweights and neck sizes of the dogs.

The purpose of the neck sizes is to get an

impression of size of collar, if radio-collaring in

the future is needed and desirable.

kg = kilogram cm = centimetre

Adult dogs*:

Dog

Sex Number of times bodyweight taken

Average bodyweight in kg.

Average neck size in cm.

Zawadi #006 F 4 20,2 36

Kimondo #040 F 2 19,6 35

Chupu Chupu #372 M 3 24,0 39

Bahati #325 M 2 24,6 39

* The dogs involved were weighed in the period 23-11-01 to 06-12-02 but not subsequently.

The average bodyweight of the adult dogs is in agreement with the range of bodyweight range

of adult dogs in the period 1995-2001, i.e. males 22-25 kg, females 18-21 kg.

23

The average neck size of the adult dogs, 39 cm for the males and 36 cm for the females, is one

cm less than the adult dogs in 1999 i.e. 40 cm for the males and 37 cm for the females.

Litters:

Malaika litter (born 07-07-01) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm.

Average bodyweight in kg.

Average neck size in cm.

23-11-01 4 9,9 9,3

15-12-01 5 13,0 30 11,9 28

15-03-02 8 17,0 34 16,0 33

06-12-02 17 22,1 37 19,2 34

05-12-03 29 21,4 36 19,2 34

03-12-04 41 19,9 36 17,7 35

December 2004 the average bodyweight of the males was approximately 3 kg, and for the

females 2 kg, too low. At all other dates the dogs of this litter had average bodyweights based

on the bodyweights of the dogs in the period 1995-2001 at the same age which are taken as

the normal value.

The Malaika litter is used as point of reference for all the litters below. As diagram 1

shows these pups grew up in a consistent way due the fact that they were on their own after

the mother died and thus not living in a pack. In this way this litter is a good point of

reference to compare other litters but one has to keep in mind that pups being raised in a pack

might deviate in bodyweight depending on the composition of the pack and the social

behaviour within the pack. However, this deviation should not be more than a couple of

kilograms and should be made up at the time the dogs are 17-20 months old when they would

normally have been expected to have reached average adult bodyweight.

Diagram 1: Growth rate Malaika litter

0

5

10

15

20

25

30

35

4 5 8 17 29

age in months

Bodyweight in kg

Male Malaika litter

Female Malaika litter

Lineair (Male Malaika litter)

Lineair (Female Malaika litter)

24

Kimondo I litter (born 18-04-02) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm.

Average bodyweight in kg.

Average neck size in cm.

06-12-02 9 15,2 30 15,8 30

05-12-03 21 16,4 32 15,9 32

03-12-04 33 14,5 31 15,3 34

December 2003 and 2004 the average bodyweight for the males was 6-8 kg and for the

females 4 kg too low.

Kimondo II litter (born 31-05-03) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm.

Average bodyweight in kg.

Average neck size in cm.

05-12-03 6 9,4 25 8,4 25

25-11-05 30 18,5 36

Average bodyweight approximately 4 kg too low for males and females.

Kimondo IV litter (born 10-04-05) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm

Average bodyweight in kg.

Average neck size in cm.

25-11-05 7 10,5 27 9,5 24

Zawadi I litter (born 11-05-04) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm.

Average bodyweight in kg.

Average neck size in cm.

04-12-04 7 7,4 24

25-11-05 19 14,5 33

Compared with the Malaika litter, average bodyweight for the males at 5 months 13 kg, the

average bodyweight of the pups of the Zawadi litter was far too low December 2004. One

year later the dogs still had not made up arrears.

Zawadi II litter (born 22-05-05) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm.

Average bodyweight in kg.

Average neck size in cm.

25-11-05 6 11,1* 28*

* Only four out of five pups were measured as one pup was insufficiently sedated.

Mrefu I litter (born 27-03-05) Date Age in

months Male Female

Average bodyweight in kg.

Average neck size in cm.

Average bodyweight in kg.

Average neck size in cm.

25-11-05 8 12,9 30 13,2 31

25

In general one might conclude that in December 2004 the bodyweights were too low.

However, most dogs did not appear to be underweight. It was observed that some

subordinates dogs did not look thinner than others of higher rank. This might have caused the

people in charge to miss the physical signs/symptoms that all the dogs were underweight,

most likely as a result of underfeeding. Measures were taken immediately as described in

chapter Nutrition.

VETERINARY

Preventative Medicine

Preventative medicine is the most important part of the work and takes the most time. It is far

better to prevent diseases than to cure diseased animals, particularly in the given situation

whereby many animals are kept in

relatively close confinement and

contagious diseases will spread easily.

Sadly enough the importance of the

preventative medicine was heavily

emphasized by the canine distemper

outbreak December 2000. As it was

impossible to combat the disease

successfully with medications available

nowadays, the only option left was the

prevention of the disease by vaccination

to protect the dogs in our breeding

program.

Next to keeping the dogs in the best possible physical shape, the prevention of canine

distemper, rabies and parvovirus infections is most important. These infections being the most

threatening infectious diseases. In the past a program was developed in which the dogs were

vaccinated against these diseases. In addition, their blood was taken on a regular basis to

study the effect of the vaccinations, which made it possible to alter the vaccination schedules

according to the results. As the results for rabies and parvovirus vaccinations were satisfying

in the past, the same vaccination schedules for these diseases were continued. The canine

distemper was a completely different story; a new vaccine and schedule was used.

Vaccination policy for the different diseases and results will be presented and discussed in the

chapter on Vaccines.

Another important part of preventative medicine is the prevention of parasites, which will be

reported and discussed in the chapter on Parasites.

26

Clinical Work--Diseases

Our policy was, and remains, to interfere as little as possible in the breeding packs. Diseased

dogs were treated only when it was possible to separate them, treat them and replace them

immediately, or when pack treatment was required and possible.

Pathology

Only a few dogs died. These deaths are reported and discussed in the chapter on pathology.

Preventative Medicine

Vaccinations.

The African wild dog appears to be extremely susceptible to diseases like canine distemper

virus, parvoviral disease and rabies. Little is known about the effectiveness of vaccinating

wild dogs against these diseases. The purpose of vaccinating our captive wild dogs is firstly to

attempt to protect them from the above-mentioned diseases and secondly to study the

effectiveness of the vaccinations. As a common rule, only inactivated (i.e. killed) vaccines

were used for two reasons: first, to prevent the introduction and spread of viruses in the

environment, and secondly, to prevent any ill side effects on the dogs.

Canine Distemper Virus (CDV) – (See Appendix II and III) Introduction

From 1995 to 2001 the CDV-Iscom vaccine, developed and donated by the Institute of

Virology, Erasmus University, Rotterdam, was used. At the first real challenge however it

appeared that this vaccine did not provided

enough resistance. As announced in the

1995-2001 report, (chapter Steps

Taken/Considered), the choice fell upon

Purevax™ (Merial). According to the

manufacturer, Purevax™ is a lyophilized

vaccine of a recombinant canarypox vector

expressing the HA and F glycoprotein’s of

canine distemper virus. More popular: a

“canarypox-vectored canine distemper

vaccine”; i.e. a distemper vaccine where

genetic information of the distemper virus

is built into the canarypox virus. The virus may multiply after injection and enhances the

building up of immunity this way. The vaccine is developed for use in ferrets and tested in

ferrets, domesticated dogs, and lesser pandas. Determination of the serum antibodies in

response to the vaccination will be continued. However, it is a live attenuated vaccine, not an

27

inactivated one, which is in contradiction with our vaccination policy not to use live

attenuated vaccines. But the vaccine-virus multiplies in the cell it enters only. It is not

excreted from that cell and is not passed on to the next cell. This way it is impossible for the

vaccine-virus to spread itself in the environment.

Immunity is developed in two different ways. There is humoral immunity and cellular

immunity. The humoral immunity is the response of B-lymphocytes to an invading infectious

agent in the form of antibodies that are produced by plasma cells that originate from the B-

lymphocytes. The cellular immunity is not the response in the form of antibodies but the

effects or reactions of different cells itself. It is the response in the form of T-lymphocytes.

What has been done in the past in the Mkomazi dogs was the study of humoral immunity, i.e.

the study of the antibodies. However, in some cases the study of cellular immunity is more

important than the study of humoral immunity as some intracellular parasites like the

intracellular life because of the protection it offers. In the cell it is impossible for humoral

antibodies to reach them (ROITT & DELVES, 2001). One of these cases is the group of

morbilli viruses and canine distemper virus is a morbilli virus.

Therefore, next to the study of humoral immunity, a study of cellular immunity (cell

mediated immunity) had to be set up. We are very happy that this will be undertaken at the

Institute of Virology, Erasmus University, Rotterdam, as well.

To increase the number of pups participating in the cell

mediated immunity research, i.e. to come to results more

quickly, we were very happy that Artis, Amsterdam Zoo, the

Netherlands, agreed to have their pups participating in the

research program. This happened with two litters. One litter in

2001 eight pups and the other litter in 2002 twelve pups. Also a

comparison was made between pups vaccinated with

Purevax™ and pups vaccinated with CDV-Iscom. Separately

will be reported over the results of the Artis pups.

However, due to the unavailability of the Purevax™ for us, we

had to decide December 2004 either to continue with the CDV-

Iscom vaccine or stop vaccinating against CDV until the

Purevax™ would be available for us again. As, after

vaccinating with CDV-Iscom vaccine, the canine distemper

outbreak in December 2000 ran a devastating course it was decided to stop vaccinating

against CDV temporarily.

Method

The following vaccination schedule was adopted for the use of Purevax™: The first year two

vaccinations with one month in between followed by annual booster vaccinations.

For the purpose of antibody testing (humoral immunity) and cell mediated immunity study

EDTA blood had to be collected at the time of the first vaccination, at the time of the second

vaccination (one month after the first vaccination) and three months and one year after the

second vaccination. For the cell-mediated immunity study EDTA blood should preferably not

be older than 72 hours at the time of arrival at the University. Thanks to a good cooperation of

all the people involved, in Mkomazi as well as in Rotterdam, this succeeded on every

28

occasion. This procedure was undertaken for the first litter of pups born after the distemper

disaster (Malaika litter 07-07-2001), seven pups in all. The same happened to the new dogs

#006 and #040 and the last two distemper epidemic survivors #372 and #325. Only #006

followed the whole procedure but quitted the program after her transfer to the Lendanai boma

to start her own pack, i.e. she received her vaccinations but was not sedated anymore for

blood samples. The same happened to #040 March 2002 as she was pregnant at that time and

the same applies to her mate Alpha male #372. Unfortunately #325 died March 2002, so we

could not follow up his antibody history anymore.

The pups of the second litter born after the distemper epidemic, Kimondo I litter, were

vaccinated according a slightly different schedule. They were vaccinated three times with

three weeks between the first and second and between the second and third vaccination of

Purevax™. Blood samples were taken only at the time of the first vaccination and one year

later.

The next litter, Kimondo II litter, was not vaccinated the first year to act as a control group.

Results humoral immunity.

1. Pre-vaccination titres to CDV: Only three dogs had a pre-

vaccination titre, #104 a titre level of 80 and #092 and

#853 a titre level of 20. All three were the only dog in their

litter with a titre.

2. Post-vaccination titres: Malaika litter: One month after

the first vaccination there were no titres, 3 months after the

second vaccination four of the seven dogs had a titre and

after one year all but one dog had a titre. Two years after

the second vaccination, with an annual booster after one

year, once again only one dog had no titre. Three years

after the second vaccination with an annual booster after

one and two years all seven dogs had a titre. Kimondo I

litter: One year after three vaccinations all four dogs had a titre. After two years with

an annual booster after one year, both dogs tested had a titre.

3. The two new, adult, arrivals #006 and #040 had no titre at the time of their first

vaccination and again no titre one month after the first vaccination. Three months after

the second vaccination and one year after the second vaccination #006 still had no

titer.

4. In the control, non vaccinated, group ( Kimondo litter II) 7 months after birth one dog

showed a titre of 20. The other dogs showed no antibodies.

Table 2: Virus Neutralizing antibody Titres in African Wild Dogs during a vaccination trial

with Purevax™ vaccine Date Time-lapse between

2nd or 3rd vaccination And date of testing

Number of dogs VNT-titre >20

% Number of dogs VNT-titre <20

% Total number of dogs

15-03-02* 3 months 4 50 4 50 8

06-12-02* 1 year 6 75 2 25 8

05-12-03* 2 years 6 85 1 15 7

05-12-03** 1 year 4 100 0 0 4

03-12-04* 3 years 7 100 0 0 7

03-12-04** 2 years 2 100 0 0 2

29

* Completed 2 initial vaccinations

**Completed 3 initial vaccinations

Results cellular immunity

Up until now (November 2004), there has been a technical problem at the University with the

growth of white blood cells, lymphocytes, required for the study of cell mediated immunity.

This problem has to be solved first before it is possible to get results.

We hope that the cell mediated immunity study will provide an answer to the question why

the canine distemper epidemic ran such a disastrous course.

Discussion/Conclusion

The number of dogs in the study is low. It appeared that dogs initially vaccinated three times,

with three weeks between vaccinations, show a slightly better response than dogs that started

initially with two vaccinations: 100% as against 75%. However, after two and three years

respectively there was no difference, 100% both.

As appendix II shows titres in the years 2002-2004

are considerably higher than titres in the years 1995-

2001 after vaccination with CDV-Iscom (VISEE et

al, 2001).The titre levels ranged from <20 to 80 in the

years 1995-2001, compared with a range of titres

from <20 to 640 in the years 2002-2004.

Although CDV-Iscom vaccine is known for its low

titres, >=20, but to provide good immunity in seals

(Van de Bildt, pers.com.), the vaccine obviously

failed to provide sufficient immunity in our African

wild dogs in 2000 (VISEE et al, 2001). The level of antibodies, after vaccination with

Purevax™, suggests that vaccination with Purevax™ provides higher antibody titres and thus

may provide a better protection against canine distemper virus.

At this moment to protect our African wild dogs, we prefer to use Purevax™ vaccine

following a vaccination schedule which provides three initial vaccinations with three weeks in

between followed by an annual booster vaccination. This schedule is advised for ferrets by the

manufacturer Merial.

Rabies (See Appendix IV).

Introduction/Method

Initially, in 1995, rabies vaccination was started with a single vaccination. Due to a

complete lack of sero-conversion the vaccination schedule was changed. From 1996

onward the dogs received three vaccinations in the first year: a second and a third

vaccination was given one and five months after the first vaccination respectively, followed

by an annual booster vaccination. This schedule proved to be successful as reported in the

1995-2001 report. It was only natural to carry on according this schedule. Vaccine used in the

year 2001-2005 was Vanguard® *.

30

Results

1. Single vaccination: December 15th 2001 three weeks after a single vaccination only

one dog of the Malaika litter (#269) developed what is considered to be sufficient

antibodies: >0.5 I.U./ml.

2. Two vaccinations: March 15th 2002 four months after the first vaccination not a single

dog of the Malaika litter showed sufficient antibodies.

3. Three vaccinations: One year after the first vaccination only one dog of the Malaika

litter showed sufficient antibodies, once again #269. However all four dogs of the

Kimondo I litter showed sufficient antibodies.

4. Three vaccination + a single annual booster vaccination: Four of the seven dogs of the

Malaika litter showed sufficient antibodies.

5. Three vaccination + a single annual booster vaccination after one and two years: Six

of the seven dogs of the Malaika litter showed sufficient antibodies.

6. The two new, adult, arrivals #006 and #040 showed no sufficient antibodies

independent of the number vaccinations.

Table 3: Rabies antibody titres in African wild dogs after vaccination with inactivated

commercial rabies vaccine: Vanguard® R (Pfizer) in the period 2001-2005.

* a Rabies specific VNA titre of >0.5 International Units (I.U./ml) is considered to be

the minimum titre likely to provide protection against challenge.

** received booster vaccination 9 months after third vaccination.

*** received booster vaccination 9 and 21 months after third vaccination.

Table 4: Rabies antibody titres in African wild dogs after vaccination with inactivated

commercial rabies vaccines: Dohyrab® (Solvay Duphar) in 1995&1997 and Rabdomun®

(Schering-Plough) from1998 to 2001 (from VISEE et al, 2001) in the period 1997-2001.

* a Rabies specific VNA titre of >0.5 International Units (I.U./ml) is considered to be

the minimum titre likely to provide protection against challenge.

** received booster vaccination at the time of blood sampling.

Discussion/Conclusion

Although numbers of dogs are low, it is interesting to compare the results of the titres after

three vaccinations with the results over the period 1995-2001 (Visee et al, 2001).

Months after third rabies vaccination

Number of dogs >0.5 I.U./ml*

% Number of dogs <0.5 I.U./ml*

% Total number of dogs

5-6 11 90,9 1 9,1 4

9 1 14 7 86 8

18 2 100 0 0 2

21** 4 57 3 43 7

33*** 6 86 1 14 7

Months after third rabies vaccination

Number of dogs >0.5 I.U./ml*

% Number of dogs <0.5 I.U./ml*

% Total number of dogs

1 12 92 1 8 13

5 13 100 0 0 13

12** 17 85 3 15 20

24 6 60 4 40 10

31

In that period 5 months after the third vaccination 100% of the dogs tested had a titre >0.5

I.U. (table 4). In the period 2001-2005 90,9% of the dogs tested had a titre >0,5 I.U. (table 3).

However, in the latter case it concerned 11 dogs against 4 dogs in the period 1997-2001.

Also, 21 – 24 months after the third vaccination, with an annual booster vaccination in

between, percentages are in the same order: 60% titre >0.5 I.U. in 1997-2001 and 57% in the

period 2001-2005.

Not in accordance with these results were the titres 9-12 months after the last vaccination. In

the period 1997-2001 85% of the dogs had a titre >0.5 I.U., in the period 2001-2005 14%, i.e.

one dog had a titre >0.5 I.U... This

concerned the Malaika litter. One

year and one booster vaccination later

the same dogs fitted into the expected

range. We have no explanation for

these, temporarily, abnormal titres.

After two annual booster vaccinations

86% of the same dogs in the Malaika

litter had a titre >0.5 I.U. which

indicates that following the

vaccination regime used a

satisfactory titre was eventually

achieved.

All in all it may be concluded that three vaccinations with an inactivated vaccine, according to

the above mentioned schedule, results in production of sufficient antibodies.

* Vanguard® Pfizer Animal Health B.V.

Parvovirus (CPV)

For vaccination the inactivated vaccine Dohyvac I-LP® (Solvay Duphar) was used for

protection against parvovirus and leptospirosis. The vaccination protocol of the manufacturer

for domesticated dogs older than 12 weeks was adopted: two vaccinations with a four-week

interval followed by an annual, single booster vaccination. No antibody testing was performed

due to positive results in the past. See Report 1995-2001.

In 2005 inactivated parvo virus vaccine was not available any more. It was decided to follow

our policy of using inactivated vaccines only. Thus the vaccination against parvo virus was

temporarily suspended except for the dogs that are not going to be used for re-introduction.

They were vaccinated with Fel-O-Vax®* i-CHP (Fort Dodge), an inactivated feline enteritis

vaccine.

* Fel-O-Vax®* i-CHP, Fort Dodge.

32

Parasites

Endoparasites

To keep healthy dogs in a breeding program in captivity an anthelmintic regime is obligatory.

Preventing worm-infection is preferable to combating the problem. Therefore twice a day the

faeces were removed from the boma’s in a very diligent way, to prevent the soil being

infested with worm-eggs as much as possible.

To check the anthelmintic status of the dogs faeces

samples were taken; once or twice a year two to five

samples per boma depending on the number of dogs in the

boma. These samples were preserved with a 5% formalin

solution before sending the samples to a laboratory.

To complete the regime the dogs received an anthelmintic,

Drontal® Dog*, on a regular basis, in a dosage as

prescribed by the manufacturer for domesticated dogs, i.e.

one tablet per 10 kg bodyweight. Drontal® Dog was

chosen as the drug of choice as it is a broad-spectrum

anthelmintic (effective against round-, tape-, and

hookworms) that needs to be administered only once and

no empty stomach required, which makes it possible to

mix the tablets with the food. As no side effects are known it may be administered to pregnant

females and pups as well.

Every two months the adult dogs received a dosage Drontal® Dog, i.e. 15 mg febantel, 14,4

mg pyrantel and 5 mg praziquantel per kg bodyweight. Pregnant females received treatment

just after mating, again 10 days later to prevent the unborn pups from being infected in the

uterus and also 10 days after giving birth. The pups received the first treatment the moment

they came out of the den and started to eat solid food, regurgitated by the pack members. At

these occasions the tablets were put in the food of the adult dogs, which made it possible to

administer an anthelmintic at an earlier age. In this way the adult dogs received some extra

treatments as well, which causes less secretion of worm eggs by the adult dogs when the pups

were at the smallest and most vulnerable.

As a result all the faeces samples proved to be negative in the years 2001-2004, which does

not mean that we exterminated all the worms but kept them at a low, apparently acceptable,

level. November 2005 a few Ancylostoma sp. eggs were found in the faeces of the dogs in the

Kisima boma.

From December 2005 onward a change was made in

the anthelmintic used. Instead of the Drontal® Dog*,

Program Plus®** is being used. It might be beneficial

to change anthelmintic every now and then to prevent

the parasites getting used to the drugs. Another

advantage was that by using Program Plus®** the

quantity of tablets that have to be administered to the

dogs can be reduced considerably as Program Plus®**

is a combination of an anthelimintic (milbemycine)

33

and an Insect Development Inhibitor (lufenuron). Thus, a tablet has to be administered only

once a month rather than twice. Dosage used as prescribed by the manufacturer for

domesticated dogs, i.e. 0.5 mg milbemycine and 10 mg per kg bodyweight.

Since December 2003 at every occasion of sedation Stronghold®*** drops were applied onto

the skin of neck of the dog in a dosage according to their bodyweight as prescribed by the

manufacturer for domesticated dogs. Stronghold® is effective against round- and tapeworms

and fleas.

*Drontal® Dog (praziquantel, pyrantelembonaat, febantel), Bayer B.V.

**Program Plus®** (milbemycine, lufenuron), Novartis Consumer Health B.V./Animal

Health

*** Stronghold® (selamectin), Pfizer Animal Health B.V.

Ectoparasites

Just as in the case of the endoparasites, keeping dogs in relatively close confinement, is bound

to be a good nurture ground for fleas and ticks. As a consequence preventive treatment has to

be enforced to keep the levels of the ectoparasites as low as possible.

On each occasion when a dog was sedated, the skin was carefully checked for parasites. There

were no signs of fungus infection or scabies. Fleas were detected occasionally. Ticks were

noticed to a somewhat bigger extent. These satisfying results are due to the following anti flea

and tick protocol.

To prevent fleas from multiplying the dogs receive every month Program®* in their food. As

Program® is not an insecticide but reduces the number of larvae by inhibiting the

development of the flea eggs, an insecticide has to be used to combat fleas. Therefore at every

sedation Advantage®*** was applied onto the neck of the dog to top up the monthly

Program® administration. December 2003 Advantage®*** was replaced by Stronghold® and

December 2005 Program® was replaced by Program Plus®** as explained in chapter on

endoparasites.

Unfortunately Program® does not affect ticks. To reduce the breeding environment for the

ticks grass in and around the boma’s is kept low with satisfactorily results up until now.

*Program® (lufenuron), Novartis Consumer Health B.V./Animal Health

**Program Plus®** (milbemycine, lufenuron), Novartis Consumer Health B.V./Animal

Health

***Advantage® (imidacloprid), Bayer B.V.

34

Clinical Work-Diseases

Clinical work

Just one exception a diseased dog had to be treated the past years. Halfway through March

2002 one of the pups of the Malaika litter, #202, was behaving in a rather sluggish way: after

resting he was the last to get up and was the first to lie down; he behaved very lethargically

and showed little interest in eating. As this coincided with the time when blood samples were

taken an examination of his blood was carried out, but this showed no abnormalities. The dog

was put on an oral course of Synulox®* and recovered quickly.

Small physical abnormalities were noticed in two dogs while sedated. The left testicle of

#269, male, was not completely descended. The testicle appeared to be just in front of the

scrotum.

The right canine tooth, lower jaw, of the female #348, appeared to protrude laterally rather

than upward.

* Synulox® (amoxicillin, clavulanic acid), Pfizer Animal Health B.V.

Anaesthesia

Over the years, the combination of medetomidine (Domitor®*) and ketamine HCL (100

mg/ml), with atipamezole (Antisedan®**) as an antidote for Domitor®*, proved to be a safe

way of sedating the dogs. From 2001 to 2005 110 sedations (see table below) have been

performed.

Table Number sedations 2001-2005 Date Number sedations

13-03-01 2

23-11-01 11

15-12-01 11

15-03-02 8

06-12-02 13

05-12-03 17

03-12-04 20

25-11-05 28

7 occasions Total 110

The drugs were administered by blowpipe,

intramuscularly, in the hindquarters. For that

purpose, the dogs were enclosed in a

passageway. The passageway was divided into

small compartments, with 2 or 3 dogs per

compartment, to avoid confusion during the

darting process.

Next quote from the Report 1995-2001 still

35

applies except for a small change.

“Over the years the adult dogs usually received 1.0 ml Domitor®* per 10 kg bodyweight,

topped up with 0.1 ml ketamine HCL per dog--independent of their bodyweight. Pups and

adolescent dogs needed a larger dosage: 1.5 ml Domitor®* per 10 kg bodyweight, topped up

with the ketamine HCL. Using the dosage of the adult dogs in the young ones meant that most

of them needed an extra administration of Domitor®*.”

From December 2002 onward the dosage of ketamine HCL was raised to 0.2 ml per dog

independent of their bodyweight. As a result no more dogs needed a ‘top up’ with Domitor®

except for dogs described in the next paragraph.

Induction time varied from 5 to 15 minutes. Some

dogs were able to remove the syringe before it

was completely emptied. Therefore, these, and

other dogs who had received too little drug due to

the mechanical failure of the syringe, received an

extra 0.5 ml of Domitor®*.

This combination Domitor®* and ketamine HCL

proved to be very satisfactory for purposes such

as taking blood samples, applying transponders,

measuring their bodyweight, etc.

The smaller dogs (<20 kg) received 1.5 ml Antisedan®**, and the larger dogs (>20kg) 2.0 ml

Antisedan®**, immediately after the veterinary procedures involving them ended--on

average 30 minutes after the administration of the Domitor®*.

Recovery time was 3 to 15 minutes, with little side effects from the ketamine HCL. The

raising of the dosage of ketamine HCL did not increase the side effects. The very slight side

effects observed were: staring into the distance and ataxia, which disappeared usually within

30 minutes and on four occasions a small seizure lasting less than a minute.

* Domitor® (medetomidine hydrochloride 1 mg/ml), Pfizer Animal Health B.V. ** Antisedan® (atipamezole hydrochloride 5 mg/ml), Pfizer Animal Health B.V.

Pathology

In the period 2001-2004 just three dogs died; two adults, #306 and # 325, and one pup. Information on #306 and the pup is extensively reported in Report 1995-2001 (VISEE et al,

2001).

The dogs #306 and #325 were two of the three survivors of the canine distemper epidemic,

which leaves #372 as the only living survivor today. #325 suddenly collapsed whilst eating on

the night of 20 March 2002 and died within five minutes. The manager of the dogs, Sangito,

was alone and too distressed to perform an autopsy. However, based on the reported

symptoms, gasping with wide-open mouth prior to collapsing, it is not unreasonable to

assume that #325 died of a heart failure, perhaps in part induced by his earlier traumatic

experiences. Sadly with him the last blood of the original Lendanai dogs disappeared.

36

Acknowledgments

My sincere gratitude for their help and support goes to:

Ministry of Natural Resources & Tourism, Government of the United Republic of

Tanzania.

Department of Wildlife, Tanzania.

Tanzania Wildlife Research Institute (TAWIRI).

George and Natasha Duffield for funding part of the program.

Duncan Forbes, Ray Rowe Trust for Animals, for funding part of the program.

Prof. A.D.M.E. Osterhaus, Marco van de Bildt and Joost Philippa, Institute of Virology,

Erasmus University, Rotterdam, for performing the antibodies testing and most of all their

invaluable advice.

Roger Burrows, for giving us invaluable insights into the behaviour of The African Wild Dog

and his comments on this report.

Mark de Boer, Diergaarde Blijdorp, Rotterdam, the Netherlands for examining the faeces

samples.

Gilbertson & Page, England, for generously supplying dry dog food, GILPA “Value

Complete”.

Pfizer, the Netherlands for generously supplying Domitor®, Antisedan®, Vanguard®,

Synulox® and Stronghold®.

Bayer, the Netherlands for generously supplying Drontal ® Dog and Advantage®.

Novartis, the Netherlands for generously supplying Program®.

Virbac, the Netherlands for generously supplying transponders.

I.U.C.N. Canid Specialist Group.

Tony Fitzjohn, field-director, for his invaluable support and help with the fieldwork

Lucy Fitzjohn, for her support and preparing annually notes, which made Sangito’s notes

accessible for me.

Sangito Lema, keeper of the dogs, for his help with the fieldwork and his valuable notes

Ayubu Mbise, assistant keeper of the dogs, for his help with the fieldwork.

Dierenkliniek Visee, Netherlands, for generously supplying medical necessities.

Aart Visee, for supplying the photos used in this report.

Sylvia Kappe†, for the final layout.

37

Summary

In 1995, the George Adamson Wildlife Preservation Trusts, and The African Wild Dog

Foundation, through the Ministry of Natural Resources and Tourism, and the Department of

Wildlife, Tanzania, started a breeding program for the African Wild Dog in Mkomazi Game

Reserve, Tanzania. This Report reports over the years 2001-2004.

In the first section, the development of the breeding packs is described, including the zoo

technical aspects. The second section deals with the veterinary aspects.

In order to protect the dogs from infectious diseases they were vaccinated against distemper,

rabies and parvo-viral disease. To establish the effectiveness of the vaccinations, testing for

antibodies was performed.

Two different litters of dogs were vaccinated against distemper (Purevax®) according to two,

different vaccination schedules. One litter received two vaccinations (one month in between)

followed by an annual booster vaccination. The other litter received three vaccinations (three

weeks between vaccinations) followed by an annual booster vaccination. In both cases

satisfactory levels of antibodies were produced

Three rabies vaccinations in the first year--one month between the first and second, and five

months between the second and the third, followed by an annual booster, gave satisfactory

levels of antibodies.

38

References

BURROWS, R. (1992): Wild Dogs of the Serengeti Part 2. Miombo, 8, 4-5.

BURROWS, R. (1995): Demographic changes and social consequences in wild dogs 1964-

1992. In: Serengeti II : research, management, and conservation of an ecosystem (Ed.

A.R.E.Sinclair & P.Arcese) Chicago :Chicago University Press.

BURROWS, R., HOFER, H. & M.L. EAST (1995): Population dynamics, intervention and

survival in African wild dogs (Lycaon pictus). Proc. R. Soc. Lond. B 262, 235-245.

BURROWS, R. (2004): Pack cohesion in the African wild dog (Lycaon pictus) and a "young-

male first" protocol in the acquisition of dominance. In : Supplements to Ethology; Advances

in Ethology 38 :124.

CARBONE, C., DU TOIT, J.T. and I.J. Gordon (1997): Feeding success in African wild

dogs: Does kleptoparasitism by spotted hyaenas influence hunting-group size? J.Anim.Ecol.

66 : 318-326.

FANSHAWE, J.H. and C.D. Fitzgibbon (1993): Factors influencing the hunting success of an

African wild dog pack. Animal Behaviour 45, 479-490.

FRAME, L.H., MALCOLM, J.R., FRAME, G.W. and H. van LAWICK (1979): Social

organization of African wild dogs (Lycaon pictus) on the Serengeti plains. Zeitschrift für

Tierpsychologie: 50, 225-249.

FRAME, G.W. and L.H. FRAME (1981): Swift and Enduring: Cheetahs and Wild Dogs of

the Serengeti. New York: E.P. Dutton.

GIRMAN 1993. Molecular Genetic and Morphological Analyses of the African wild dog

(Lycaon pictus). Journal of Heredity. 84, 450-459.

GIRMAN, D.J., MILLS, M.G.L., GEFFEN, E. & R.K. WAYNE (1997): A molecular genetic

analysis of social structure, dispersal, and interpack relationships of the African wild dog

(Lycaon pictus). Behav. Ecol. Sociobiol. 40, 187-198.

MACNUTT, J.W. (1996) : Adoption in African wild dogs, Lycaon pictus. J. Zool. Lond. 240,

163-173.

MALCOLM, J.R. (1979): Social organisation and communal rearing in African wild dogs.

PhD Thesis, Harvard University.

MALCOLM, J.R.and K. Marten (1982): Natural selection and the communal rearing of pups

in African wild dogs (Lycon pictus). Behavioural Ecology & Sociobiology 10: 1-13.

ROITT, I.M. and P.J. DELVES (2001): Roitt’s Essential Immunology. Blackwell Science

Ltd..

SCHALLER, G. B. (1972): The Serengeti Lion. The Chicago University Press.

VISEE, A.M., FITZJOHN, A.R., FITZJOHN, L. and S. LEMA (2001): African Wild Dog

(Lycaon pictus) Breeding program, Mkomazi Game Reserve, Tanzania, Report 1995-2001.

Unpublished. George Adamson Wildlife Preservation Trusts, The African Wild Dog

Foundation.

39

Appendix I

Date

23-11-01

15-12-01

15-03-02

06-12-02

05-12-03

03-12-04

25-11-05

Identification

Sex

of Birth

kg

ns

kg

ns

kg

ns

kg

ns

kg

ns

kg

ns

kg

ns

Zawadi

006

F20,6

38

20,1

37

20,2

35

19,8

35

Kimondo

040

F20,0

35

19,3

35

Bahati

325

M04-09-97

24,3

39

24,9

39

Chupu Chupu

372

M08-01-99

23,0

38

24,3

39

24,8

40

Malaika

448

F07-07-01

9,4

12,1

28

15,8

32

19,1

34

18,6

34

litter

464

F07-07-01

9,3

11,7

28

16,2

34

19,3

35

19,7

35

202

M07-07-01

9,0

11,9

26

14,7

31

21,0

37

22,0

36

18,4

35

251

M07-07-01

9,1

13,3

29

16,5

34

21,6

37

22,3

37

19,9

36

269

M07-07-01

10,0

13,0

31

18,0

34

21,6

36

20,2

35

368

M07-07-01

10,8

13,7

32

17,9

35

24,4

39

21,8

37

389

M07-07-01

10,4

31

17,6

34

21,8

36

20,6

35

Kimondo I

104

F18-04-02

17,1

31

17,0

32

16,2

33

litter

348

F18-04-02

14,5

30

14,7

31

14,4

32

651

M18-04-02

14,1

29

15,5

31

14,1

30

730

M18-04-02

16,2

32

17,3

34

14,8

32

Kimondo II

001

F31-05-03

8,5

25

litter

031

F31-05-03

8,4

25

092

M31-05-03

10,8

27

18,9

38

187

M31-05-03

9,7

24

19,4

36

212

M31-05-03

9,1

26

18,6

36

393

M31-05-03

8,0

24

17,0

35

Zawadi I

666

M11-05-04

6,8

24

14,2

32

litter

671

M11-05-04

7,3

25

14,2

34

732

M11-05-04

6,5

22

12,9

32

853

M11-05-04

7,9

24

14,8

32

860

M11-05-04

7,4

25

15,8

34

887

M11-05-04

8,0

25

14,7

32

926

M11-05-04

8,6

25

15,9

33

939

M11-05-04

7,0

25

13,7

32

949

M11-05-04

7,0

23

14,5

33

Zawadi II

519

M22-05-05

12,3

29

litter

521

M22-05-05

560

M22-05-05

10,4

28

567

M22-05-05

10,5

28

Kimondo IV

572

M22-05-05

11,1

28

litter

2040

F10-04-05

10,4

28

043

M10-04-05

11,4

29

046

F10-04-05

8,8

26

073

M10-04-05

9,5

26

Mrefu I

444

F27-03-05

13,2

32

litter

481

M27-03-05

12,8

31

557

M27-03-05

13,6

31

586

M27-03-05

13,9

31

643

M27-03-05

11,5

29

Bodyweight in kilograms (kg) Necksize in centimeters (ns)

40

Appendix IIDate

Identification SexZawadi 006 F V <20 V <10 <10 V T10 VKimondo 040 F V <20 V <10 V VBahati 325 M V 80 V 20 died 20-03-02Chupu Chupu 372 M V 160 V 80 V 160 VMalaika 448 F V <20 V <10 20 V 20 V 320 1280litter 464 F V <20 V <10 40 V T10 V 160 640

202 M V <20 V <10 T10 V 20 V T10 160251 M V <20 V <10 80 V 40 V 40 640269 M V <20 V <10 T20 V 40 V 40 320368 M V <20 V <10 160 V 80 V 160 1280389 M V <20 V <10 T20 V 40 V 640 1280

Kimondo I 104 F V 80 V V V 160 1280litter 348 F V T10 V V V 80 160

651 M V T20 V V V 20730 M V T10 V V V 160

Kimondo II 001 F T10litter 031 F <10

092 M 20 <10187 M <10 <10212 M T10 <10393 M T10

Zawadi I 666 M <10litter 671 M 10

732 M <`10853 M 20860 M <10887 M T926 M <10939 M <10949 M <10

Zawadi II 519 M <10litter 521 M

560 M <10567 M <10572 M <10

Kimondo IV 039 Flitter 2040 F <10

043 M046 F <10073 M <10

Mrefu I 444 F <10litter 481 M <10

557 M <10586 M <10643 M <10

Distemper antibody levels (VNT) + Vaccination dates Mkomazi23-11-01 15-12-01 15-03-02 06-12-02 29-12-02 21-01-03 05-12-03 03-12-04 25-11-05VNT titer VNT titer VNT titer VNT titer VNT titer VNT titer VNT titer VNT titer VNT titer

V=Vaccinated T=Toxid

41

Appendix IIIDate

Identification SexZawadi 006 F V <10 V <10 <10 V 20 VKimondo 040 F V <10 V <10 V VBahati 325 M V V >1280 died 20-03-02Chupu Chupu 372 M V V 640 V 160 VMalaika 448 F V <10 V <10 80 V 160 V 640 #litter 464 F V <10 V <10 80 V 80 V 80 #

202 M V <10 V <10 40 V 80 V <10 #251 M V <10 V <10 40 V 80 V 20 #269 M V <10 V <10 <10 V 160 V 40 #368 M V <10 V <10 160 V 160 V 40 #389 M V <10 V <10 <10 V 80 V 640 #

Kimondo I 104 F V 80 V V V 40 #litter 348 F V <10 V V V 20 #

651 M V 20 V V V 320730 M V <10 V V V 40

Kimondo II 001 F V <10litter 031 F V <10

092 M V 20 <10187 M V <10 <10212 M V <10 <10393 M V <10

Zawadi I 666 M <10litter 671 M 20

732 M <10853 M 20860 M <10887 M 40926 M <10939 M <10949 M <10

Zawadi II 519 M <10litter 521 M

560 M <10567 M <10572 M <10

Kimondo IV 039 Flitter 2040 F <10

043 M046 F <10073 M <10

Mrefu I 444 F <10litter 481 M <10

557 M <10586 M <10643 M <10

#= results not receivedV= Vaccination Purevax

Elisa titer Elisa titer Elisa titer Elisa titer Elisa titer Elisa titer Elisa titer Elisa titer Elisa titer

Distemper antibody levels (Elisa) + Vaccination dates Mkomazi23-11-01 15-12-01 15-03-02 06-12-02 29-12-02 21-01-03 05-12-03 03-12-04 25-11-05

42

Appendix IV Rabies antibody levels + Vaccination dates Mkomazi

Date 29-12-02 21-01-03 12-06-03

Identification Sex I.U. I.U. I.U. I.U. I.U. I.U. I.U.

Zawadi 006 F V <0.5 V <0.5 V <0.5 V <0.5 V

Kimondo 040 F V <0.5 V <0.5 V V V V

Bahati 325 M V >0.5 V >0.5 died 20-03-02

Chupu Chupu 372 M V <0.5 V <0.5 V >0.5 V V

Malaika 448 F V <0.5 V <0.5 V <0.5 V <0.5 V <0.5 V >0.5 V

litter 464 F V <0.5 V <0.5 V <0.5 V <0.5 V >0.5 V >0.5 V

202 M V <0.5 V <0.5 V <0.5 V <0.5 V 0.5 V <0.5 V

251 M V <0.5 V <0.5 V <0.5 V <0.5 V <0.5 V >0.5 V

269 M V <0.5 V >0.5 V <0.5 V >0.5 V >0.5 V >0.5 V

368 M V <0.5 V <0.5 V <0.5 V <0.5 V >0.5 V >0.5 V

389 M V <0.5 V <0.5 V <0.5 V <0.5 V >0.5 V >0.5 V

Kimondo I 104 F V <0.5 V V V >0.5 V >0.5 V

litter 348 F V <0.5 V V V >0.5 V >0.5 V

651 M V <0.5 V V V >0.5 V V

730 M V <0.5 V V V >0.5 V V

Kimondo II 001 F <0.5 V

litter 031 F <0.5 V

092 M <0.5 V >0,5

187 M <0.5 V >0,5

212 M <0.5 V >0,5

393 M <0.5 V

Zawadi I 666 M V V >0,5

litter 671 M V V >0,5

732 M V V

853 M V V >0,5

860 M V V >0,5

887 M V V >0,5

926 M V V <0,5

939 M V V >0,5

949 M V V

Zawadi II 519 M V <0,5

litter 521 M V

560 M V <0,5

567 M V <0,5

572 M V <0,5

Kimondo IV 039 F V

litter 2040 F V <0,5

043 M V

046 F V <0,5

073 M V <0,5

Mrefu litter I 444 F V <0,5

481 M V <0,5

557 M V <0,5

586 M V <0,5

643 M V <0,5

V = Vaccination

V

V V

V

V

V

V

V

V

V

V

V

V

V

V

V

V

V

V

V

V

V

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46

Genetic Analysis of African Wild Dogs (Lycaon pictus)

from the Mkomazi Project (1995-2004)

For: The the African Wild Dog Foundation

Att: Drs. A. Visee

By: Dr. Ir. A. Kappe

Gendika

Industrieweg 1

9641 HM Veendam

Date: 1 november 2005 (toevoeging januari 2007)

47

Introduction

The African wild dog (Lycaon pictus) is an extremely endangered species. In order to prevent the species for extinction a rehabilitation program was established by the George Adamson Wildlife Preservation Trusts and the African Wild Dog Foundation involving capture, captive breeding and re-introduction. In 1995 a captive breeding program was started by 25 wild dog pups from three locations in the Masai Steppe, Tanzania, forming three family-groups; Najo-group (n=13), Llonddirrigiss group (n=8) and Lendanai group (n=4). In this report a genetic analysis of the dogs involved in the Mkomazi Project will be presented.

Materials and methods

Genetic samples Samples were collected from 58 African wild dogs. Blood-samples from the original wild dogs were collected in 1995 and DNA was isolated by Dr. L. van de Zande, University of Groningen, the Netherlands (n=23). Blood-samples from pups born within the Mkomazi Project were collected per year and stored for analysis (n=18). In addition, 2 samples from wild caught animals were collected. See also Appendix 1 for a complete list of samples.

DNA isolation DNA was isolated from 5 ml whole blood collected in EDTA anticoagulant vacutainers. DNA was extracted by proteinase K digestion (100 μg/ml) and DNA was isolated and purified by using Standard Gendika protocol GDKiso06.

Identification of microsatellite alleles Wild dog samples were screened by the microsatellite technique. In the microsatellite technique highly variable microsatellites are used as genetic markers. Microsatellites, also known as simple sequence repeats or short tandem repeats, are genomic sequences that consist of di-nucleotide motif repeated in multiple tandem copies, in mammals CA-repeats are most common. The variation within the microsatellite loci has been shown to arise from variation in the number of repeat units (the number of CA-repeats). The variable appearances of a microsatellite are called alleles. Alleles present for a microsatellite are generated by PCR amplification. The lenght of the various alleles can be detected by cappilary gel electrophoresis in a Genetic Analyzer. By examination of a set of microsatellites a specific pattern of alleles will be produced for each individual. This specific pattern of alleles is also known as a DNA fingerprint.

Wild dog samples were screened for variation in 10 CA(n) microsatellite loci, originally isolated from a domestic dog library, combined in a Canine 10 Plex Kit (Applied Biosystems). Nine microsatellites were identified that consistently gave PCR product, were polymorphic in wild dogs (see Appendix 2).

48

Detection of microsatellite alleles from DNA was achieved by performing 35 cycles of PCR amplification in a 10 μl reaction volume using 10 ng of target DNA and 0.36 units of Taq DNA polymerase (Applied Biosystems). Alleles were detected by a Genetic Analyzer.

Data analysis The similarity, which is the average of shared alleles between two animals, was calculated as Sxy= 2nxy / (nx + ny), where nx and ny are the number of microsatellite alleles present in individual x and indiviual y, respectively, and nxy is the number of alleles shared by individuals x and y (Wetton et al., 1987). The similarity between animals is presented as a percentage. With this percentage the amount of Mean similarity within a group was estimated by averaging the similarities of all pairs of individuals that belong to the group. Mean similarity between groups A and B was estimated by averaging the similarities of all pairs of individuals of which one belonged to group A and the other to group B.

Results For all individual African wild dogs a pattern of microsatellite alleles could be scored and individual specific genotypes were identified, which means that for all animals a specific DNA fingerprint could be produced. The patterns of alleles for the nine microsatellites of all individual animals were compared and the percentage of shared alleles, the similarity Sxy, was calculated between animals, see Appendix 3. The calculated similarity Sxy is a measure for the relationship between animals. Parents share at least 50% of the alleles with their offspring, which means that the similarity between parents and offspring will be on average 0.5 or higher. Between siblings the similarity will be higher, than the similarity between unrelated animals, normally a similarity of 0.4 - 0.6 is found between siblings and a similarity of 0.0 – 0.3 is found in unrelated animals. In small populations all similarities will be higher, more alleles are shared between the animals because of the appearance of inbreeding within small populations.

Similarity in original dogs

The relationship within the three groups of original African wild dogs, Najo, Llondirrigiss and Lendanai, was in this project re-calculated and analysis of patterns yielded mean similarities of 0.55 - 0.58, which were comparable to the values found by van de Zande, 1997 (see Table 1).

49

Table 1: Similarity of alleles, Sxy (%) within and between original groups of African wild dogs

Similarity

Najo (n=13) Within group 0.58 Between groups N-Llondirrigiss 0.45 N-Lendanai 0.39 Llondirrigiss (n=6) Within group 0.58 Between groups Ll-Najo 0.45 Ll-Lendanai 0.37 Lendanai (n=4) Within group 0.55 Between groups Le-Najo 0.39 Le-Llondirrigiss 0.37

The values of mean similarity between the three groups were considerably lower than the values within the groups, which was also consistent with the data found by van de Zande.

Similarity in new dogs

In this analysis two newly introduced dogs, wild caught, were taken in to the research. Between the two new animals, 006 and 040, a high mean similarity of 0.78 was calculated (see Table 2).

Also a mean similarity between the new animals and the three original groups was calculated. These values were comparable to the values of similarity calculated between the three original groups. The value between animal 006 and the Londirrigiss group is 0.62, which is considered to be quite high for unrelated animals. Table 2: Similarity of alleles, Sxy (%) between new dogs and original groups of African wild dogs

Similarity

New dogs

006 040 006 - 0.78 040 0.78 - Najo 0.53 0.48 Llondirrigiss 0.62 0.51 Lendanai 0.29 0.33

50

Similarity in various litters In order to investigate the possibilities for the breeding program to combine pups from various litters the mean similarity of alleles between pups born within the Mkomazi Project was calculated, see table 3. In table 3a the similarity of alleles within the various litters is explained. In Table 3b the similarity between the various litters of variable years is indicated. And in this table also a comparison of animals in the litter of 2004 with newly formed packs of animals, called Sangito and Ayubu, is included. In both tables the litters are indicated by the combination of parents and the year the litter was born. See Appendix 1 for a complete list of animals.

Table 3a: Similarity of alleles, Sxy (%) within litters of African wild dog pups

Parents (female x male)

297 x 300

1997

262 x 274

1999

306 x 372

2001

040 x 372

2002

006 x 251

2004

S xy Within litter

0.63 (n=3)

0.72 (n=2)

0.69 (n=7)

0.72 (n=4)

0.73 (n=9)

Similarity between siblings is normally estimated between 0.4 – 0.6, in table 3a is quite evident that the similarity within the litter in the Mkomazi Project this similarity is slightly higher. Similarities calculated for the various litters is comparable to each other. Table 3b: Similarity of alleles, Sxy (%) between litters of African wild dog pups

297 x 300

1997

262 x 274

1999

306 x 372

2001

040 x 372

2002

006 x 251

2004

S xy Between litters

1997 - 0.48 0.43 0.45 0.47 1999 - 0.38 0.50 0.33 2001 - 0.57 0.62 2002 - 0.53 2004 - S xy Between litter and packs

Sangito 0.60 Ayubu 0.57

51

The similarity between the litters is considerably lower than the calculated similarity within the litters. All similarities between the litters are in the same order as the similariy calculated within the litters of the original dogs. The comparison of the litter of 2001 (306 (F) x 372 (M)) and the litters of 2002 (040 (F) x 372 (M)) and of 2004 (006 (F) x 251 (M)) show a slightly higher similarity. The newly formed packs are animals which should form a new breeding group. Two packs of animals were formed; Sangito (animal 448, 464, 651and 730) and Ayubu (animal 269, 368, 389, 104, 348).

Similarity between the newly formed packs and the litter from 2004 (006 (F) x 251 (M)) was calculated as 0.60 for the Sangito pack and 0.57 for the Ayubu pack, respectively, which is slightly higher than the similarity between the other litters, but in the same order as the comparison of the 2004 litter and the litter of 2001, 0.62. This comparison gives also a considerable lower similarity than within litter comparison of the 2004 litter, which was 0.73.

52

Conclusion

General From the results of the genetic analysis by DNA microsatellites can be concluded that there is considerable genetic variation in the African wild dog population in the Mkomazi project. As van der Zande (1997) also concluded the genetic variation in the original African wild dog population, separated in three groups, was considerable, with a mean similar ranging from 0.55 - 0.58 for related animals and a mean similarity for unrelated animals ranging from 0.37 - 0.45. In literature a similarity of 0.0 – 0.3 is described in unrelated animals, a similarity of 0.5 or higher is found for parents and offspring and a similarity of 0.4 - 0.6 is found between siblings (Avise. 1994). Girman et al. 2001, reported quite high genetic variation in other wild dog populations in Africa as well.

The “new” dogs The similarity between two new dogs 006 and 040, result of wild caught, is quite high 0.78. As mentioned above the similarity between siblings is in the order of 0.4 – 0.6. From this result the conclusion can be that the two animals are most likely close related, and probably these animals are sisters. The observed similarity between these two new dogs and the original dogs in the Mkomazi project is ranging 0.29-0.33 for the Lendanai group, 0.48-0.53 for the Najo group and up to 0.51-0.62 with the Llondirrigiss group, which is a bit lower compared to the similarities within these groups. This implies that the “new” animals, 006 and 040, can be well bred into the original population of wild dogs from the Masai Steppe. Considering the results it is therefore recommendable to combine the two females with animals originated from or related to the Lendanai group, which shows very low similarity with both new dogs. But unfortunately these dogs did not survive the Canine Distemper-virus outbreak. The female dog 040 was bred with a male dog 372, offspring of the original dog 284 (F) from the Najo group and original dog 298 (M) from the Llondirrigiss group. The observed similarity between these two dogs is 0.50, which is high for unrelated animals. Especially when the similarity is compared to the similarities between the three original groups. A higher degree of similarity can be expected in pups of this combination. This result is showed in the comparisons of the various litters, see table 3b and also the following part.

53

Future breeding A measure of relatedness in pups born from variable combinations was made by calculating the similarity within the litters of pups born in the Mkomazi Project. With the results from these calculations a status can be revealed of the genetic variation in the pups. All full-sibs combinations were calculated and resulted in a similarity of approximately 0.70, which is higher as found in the original groups of dogs, the similarity at that point was 0.55 – 0.58. Between the litters we see a considerable amount of variation, which is shown by considerable lower similarities. Therefore concluded can be that there are good possibilities to combine pups from different litter (combinations) for future breeding. Preferable combinations for breeding are; a: pups born out of combination 262 x 274 (litter 1999) and combination 297x300 (litter

1997) can be bred with pups from all other litters, depending on the male/female ratio b: pups born out of combination 006 x 251 (litter 2004) can be bred with pups born out

of combination 040 x 372 (litter 2002). For practical reasons, pups out of the litter 2002 are used in the breeding program.

All other combinations of different litters show lower similarity with other combinations, than the similarity calculated from within litters (table 3b), which implies that the average similarity will not increase by combining the pups of various combinations/litters. Inbreeding needs to be taken into account, so combinations of pups with the same parents or grandparents needs to be avoided.

Reintroduction For breeding purposes two newly fromed packs of wild dogs Sangito and Ayubu were screened on relatedness within the packs by calculating similarity within the animals in the two packs. Both formed packs showed a similarity of approximately 0.60. This similarity is lower than calculated for pups born in the various combinations, concluded can be that the Sangito and Ayubu packs can be a good start for future breeding between the offspring of various litters. The combination of pups from the litter of 006 x 251 born in 2004 will give a comparable similarity when these pups are compared with animals in the two packs and therefore these pups can be a good addition to the formed packs of wild dogs. The observed similarities in the African wild dogs within the Mkomazi project are still at a very acceptable level, when all animals are taken into account a similarity of 0.46 for all related and unrelated dogs together can be calculated. The results in this analysis of the African Wild dog from the Mkomazi Project show very good genetic opportunities for this species.

54

Literature Avise, J.C. 1994. Molecular markers, natural history and evolution. Chapman and Hall, Inc

Girman, D.J., Vila, C., Geffen, E., Creel, S., Mills, M.G.L., McNutt, J.W., Ginsberg, J., Kat, P.W., Mamiya, K.H. and Wayne, R.K. 2001. Patterns of population subdivision, gene flow and genetic variablility in the African wild dog (Lycaon pictus). Molecular Ecology, 1703-1723. Van de Zande, L. 1997. Genetic variation within and between African Wild dog (Lycaon Pictus) groups from the Maasai steppe, Tanzania. Mkomazi project. Wetton, J.H., Carter, R.E., Parkin, D.T., and Walters, D. 1987. Demographic study of a wild house sparrow population by DNA ‘fingerprinting’. Nature, 327, 147-149.

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© The African Wild Dog Foundation 2012