Hereditas 96: 299-305 (1982)

Electrophoretic variation in large mammals

11. The red fox, Vulpes vulpes, the stoat, Mustela erminea, the weasel,
Mustela nivalis, the pole cat, Mustela putorius, the pine marten, Martes
martes, the beech marten, Martes f o i n a , and the badger, Meles rneles
V. SIMONSEN

Institute of' Ecology and Genetics, University of Aarhus, Denmark

SIMONSEN, V . 1982. Electrophoretic variation in large mammals. 11. The red fox, Vulpes v d p e s , the
stoat, Mustela erminra, the weasel, Mustela nivalis, the pole cat, Mustela putorius, the pine marten,
Martes martes, the beech marten, Martes foina, and the badger, Meles meles. - Hereditas 96:
299-305. Lund, Sweden. ISSN 0018-0661. Received September 28, 1981

Twenty-one enzyme loci have been resolved by starch gel electrophoresis in liver and muscle tissue of
samples of 282 red foxes, 39 stoats, 13 weasels, 24 pole cats, 2 pine martens, 121 beech martens, and 5
badgers. No variation in the mobility of electrophoretic bands within the seven species of carnivores was
observed. The genetic variability is, thus, low as in several large mammals.

Vibeke Simonsen, Institute of Ecology and Genetics, University of Aarkus, N y Munkegade. DK-8WO
Arbits C , Denmark

The aim of the present work is to extend the in- than 15 km from the marking locality. The densi-
vestigation of genetic variation in carnivores and, ties of the species in Denmark a r e not known, but
in particular, to test the suggestion from previous on average 50,000 red foxes, 3,000 stoats, 2,500
studies by starch gel electrophoresis that these pole cats, 2,500 beech martens, and 1,500badgers
animals possess a low genetic variation. Low are shot every year according t o JENSEN(1976).
levels of variation have been found in natural This suggests that the population of red foxes must
populations, by BONNELL and SELANDER (1974) in be rather large to withstand a hunting pressure of
the northern elephant seal, by MCDERMID et al. 50,000 animals every year.
(1972) in the Macquarie Island elephant seal, and T h e chromosomal constitution of carnivores has
by ALLENDORF et al. (1979) in the polar bear. In been studied by several investigators, and a re-
addition, SIMONSEN (1976), MEERAKHANet al. view is given by WURSTER and BENIRSCHKE (1968).
(1973), WEIDENet al. (1974), and FISHER et al. From the carnivoran phylogeny depicted in the
(1976) observed low variation in a study of discussion part of the review it seems likely that
domestic dogs. the Canidae and Mustelidae have developed from
The investigation deals with red fox, Vulpes the Miacidae in the Eocene period and that, in the
v14/peS, stoat, Miistela nivalis, pole cat, Mustela Pleistocene, the dog and fox have developed from
putoriits, pine marten, Martcs martes, beech the Canidae. The chromosome number, location
marten, Murtes f o i n a , and badger, Meles rneles. of the centromere, and the nombre fondamental
All of them are carnivores and widespread in (NF) are listed in Table 1 for the dog, the polar
Europe, the species live as single individuals or in bear, and the species investigated in this study.
small families, and they breed once a year. Several According to WURSTER and BENIRSCHKE (1968) the
migration studies have been carried out on the red N F is remarkably constant within the families,
fox, for example by JENSEN(1973), who did a except for the Canidae and Mustelidae. WURSTER
marking-recovery experiment with 460 cubs. The and BENIRSCHKE (1968) state that much of the var-
red fox seems to migrate mostly in the first year of iation in the number of chromosomes among the
life, and can migrate up to 100 km. Males seem to nine species of Carnivora can b e explained by
migrate further than females, but 85 7% of the fe- Robertsonian mechanisms. Anyhow, the rather
males and 75 9% of the males d o not migrate more great similarity among the karyotypes of the
300 V. SIMONSEN Hereditas 96 (1982)

Table 1 . Chromosome number in total, divided into meta- and submetacentric chromosomes and telo- and subtelocentric chromo-
somes, and the NF value for nine species of carnivores according to WURSTER and BENIRSCHKE (I-)

Meta- and Telo- and

Family and Chromosome submetacentric subtelocentric NF
species number chromosomes chromosomes

Family Canidae
Subfamily Caninae
Canis familiaris, 78 0 76 80
domestic dog
Vulpes vulpes, 34-38a 32 4? 68-76
red fox
Family Ursidae
Thalarctos maritimus 74 12 60 88
polar bear
Family Mustelidae
Subfamily Mustelinae
Mustela erminea , 44b 16 26 62
stoat
Mustela nivalis, 42c - - -
weasel
Mustela putorius, 40 28 10 70
pole cat
Martes martes, 38 -
pine marten
Martes foina , 38
beech marten
Subfamily Melinae
Meles meles , 44 26 16 72
badger

a BUCKTON and CUNNINGHAM (1971) say that the chromosome number vanes from 35 to 39
According to MANDAHL and FREDGA (1980)
C According to FREDGA and MANDAHL (1973)

species might suggest that the level of genetic var- silver foxes and have found three loci to be poly-
iation among the carnivores is low. morphic among the silver foxes, but the authors
Previous studies on carnivore hemoglobin have d o not record an estimate of the heterozygosity.
shown that there is no electrophoretic variation The observations support the model of low genetic
among the families Canidae, Procynoidae, Ursi- variability among carnivores, when studied with
dae, Mustelidae, Otariidae, and Phocidae, as re- starch gel electrophoresis.
ported by SEAL(1969), but by use of micro-
complement fixation procedure SARICH (1972) has
been able to separate the hemoglobins from the six
mentioned carnivore families. LEMUXand KENYON
(1975) have, by use of immunoelectrophoresis,
Materials and methods
shown that the subfamilies Mustelinae, Melinae, The animals used were mainly the results of hunt-
and Lutrinae have probably shared a common ing from different localities in Denmark. The
ancestor for some time after the divergence of the number of individuals in each species and the sex
line leading to the modern Mephitinae, which of the individuals, when known, are listed in Table
supports the model for evolution of the carnivores 2.
that is based on karyotypes, as described by WUR- Muscle and liver tissue samples were removed
STER and BENIRSCHKE (1968). SEROVet al. (1976), from the animals as soon as possible, and stored at
SEROV and ZAKIJAN (1977), and SEROV et al. (1978) -70°C until use. The samples were analysed by
have studied allelic expression in intergeneric fox starch gel electrophoresis for 15 enzymes; refer-
hybrids between arctic and silver foxes. SEROV et ences for the buffer systems, and staining proce-
al. (1976) have studied 33 loci in the arctic and dures are listed in Table 3.
Hereditas 96 (1982) ELECTROPHORETICVARIATION I N LARGE MAMMALS. 11. 301

Table 2. Number and sex of the seven carnivore species used for this study

Sex Species

Red Pole Pine Beech

fox Stoat Weasel cat marten marten Badger

Male 143 19 a 19 I 51 I
Female 139 20 5 5 1 51 4
Unknown 19

Total 282 39 13 24 2 I21 5

Table 3. Enzymes analysed, references for buffer systems, and staining procedures

References
Enzyme Abbre-
viation Buffer system Staining procedure
~~

Acid phosphatase ACP 0.1 M phosphate pH = 7.0 HARRIS and HOPKINSON1976

Adenosine deaminase ADA SPENCER et al. 1964 SPENCER et al. 1964
Adenylate kinase AK FILDES and HARRIS 1966 FILDES and HARRIS 1%6
Catalase CAT HARRIS and HOPKINSON 1976 S H A W and KOEN 1%8
N ADH-diaphorase DIA As CAT GIBLETT 1%9
Esterase EST POULIK 1957, modified; SIMONSEN and FRYDENBERG 1972;
or HARRIS and HOPKINSON 1976 or HOPKINSON et al. 1974
Glucose-6-phosphate dehydrogenase G6PD As ACP GIBLETT 1%9
Glutamic-oxaloacetic transaminase GOT HARRISand HOPKINSON 1976; SCHWARTZ et al. 1%3
or as EST
Glutamic-pyruvic transaminase GFT HARRISand HOPKINSON 1976 HARRIS and HOPKINSON 1976
Isocitrate dehydrogenase 1CD HARRISand HOPKINSON 1976 HARRIS and HOPKINSON 1976
Lactate dehydrogenase LDH FlLDES and HARRIS 1966 HYLDGAARD-JENSEN 1%7
Malate dehydrogenase MDH FlLDES and HARRIS 1%6 As LDH with males as substrate
Peptidase PEP FILDES and HARRIS 1966 With leucyl-glycyl-glycin as substrate
or HARRIS and HOPKINSON 1976
6-phospho-gluconate dehydrogenase PGD As ACP GIBLETT 1%9
Phosphoglycerate kinase PGK As GPT HARRIS and HOPKINSON 1976

Results strongest activity. Only one band was found, and

no variation within the species was observed.
For the species investigated the results of the en-
zyme analyses are presented below. Diaphorase (DIA). - Liver tissue expressed the
strongest activity, and mostly one or two bands
Acid phosphatase (ACP). - Liver tissue exhi- were revealed. No genetic variation within the
bited the strongest activity. No variation among species was observed.
the individuals within the species was obtained.
The zymogram consisted of one band. Esterase (EST). - Two zones of activity were
found in liver tissue, corresponding to B2and B3 in
Adenosine deaminase (ADA). - Again liver human (HARRIS and HOPKINSON 1976). The same
tissue possessed the strongest activity. Each zones of activity were seen in muscle tissue, but
species had either one band or three bands. No with less activity. A staining procedure with
variation within the species was found. 4MUB-acetate as described by HOPKINSON et al.
(1974) did not reveal additional bands. No var-
A d e n y h t e kinase (AK). - Both liver and muscle iation within the species were observed.
tissues expressed activity, one heavy band and
one to two weaker bands. N o variation within the Glucose-6-phosphate dehydrogenase (G6PD). -
species was seen. Muscle tissue exhibited one band, whereas liver
tissue showed a rather diffuse zone of activity. No
Catalase (CAT). - Liver tissue revealed the variation within the species was detected.
302 v. SIMONSEN Hereditos % (1982)

Beech Marten
Stoat
L Weasel

Red Fox
Badger

I I I I 1 I
0.0 0.5 1.0
GENETIC SIMILARITY
Fig. 1. The genetic similarity (I) among seven species of carnivores, based on twenty-one enzyme loci.

Glutamic-oxaloacetic transuminuse (GOT). - Discussion

Both liver and muscle tissue expressed two bands.
No variation within the species was found. In Table 4 the variation in the enzymes studied
among the seven species of carnivores is listed
Glutamic-pyruvic transaminuse (GPT). - Only together with the designation of the loci and
muscle tissue expressed the activity of the en- alleles. None of the loci show polymorphism in
zyme. One main zone of the activity and addi- either species. This observation is in accordance
tional weaker zones were observed. N o variation with the results found for several species of large
within the species was revealed. mammals, as mentioned in the introduction. A
Isocitrate dehydrogenase (ICD). - One band was similar result is found for man and apes, as shown
seen in liver tissue, whereas two bands were found by BRUCE and AYALA (1979). BoNNELLand SELANDER
in muscle tissue. No variation within the species (1974) explain the lack of genetic variation in ele-
was observed. phant seals as a result of a population bottle-neck
last century. ALLENDORF et al. (1979) offer several
Lactate dehydrogenase (LDH). - As expected, alternatives for explaining the low level of hetero-
both liver and muscle tissue expressed a fiveband- zygosity in polar bears besides the effect of a
ed zymogram. N o variation within the species was population bottle-neck. Anyhow, this study
revealed. supports the hypothesis that large mammals have
Mulute dehydrogenase (MDH). - Both liver and nearly no genetic variation when enzymes a r e
muscle tissue revealed two zones of activity, each studied with starch gel electrophoresis.
with a heavily stained band, and one to two addi- The values for genetic identity and distance
tional weaker bands. No variation within the according to NEI (1972) a r e listed in Table 5 . A
species was found. pair-group clustering procedure is carried out with
unweighted arithmetic averages, and the result is
Pepriduse (PEP). - Both muscle and liver tissue depicted in Fig. 1. It is not possible to distinguish
expressed the same zymogram, presumably corre- between the pine and beech marten by this meth-
sponding to PEP B in human (HARRIS and HOPKIN-od, but the grouping of the species is in accord-
SON 1976). ance with the classification in subfamilies, by
6-Phosphogluconate drhydrogenuso (PGD). - morphological characters and by karyotypes.
Activity was expressed in both tissues, but muscle All in all, the seven carnivore species have no
tissue gave one distinct band. N o variation within genetic variation within the species, and the
the species was seen. grouping representing the electrophoretic var-
iation among the species is in agreement with the
Phosphoglycerate kinase (PGK). - Activity was expectations based on morphological characters
seen in both tissues, but again the muscle tissue and karyotypes.
exhibited two distinct bands. No variation within
the species was found.
 Hereditas 96 (1982) ELECTROPHORETIC VARIATION INLARGE MAMMALS. 11. 303

Table 4. Enzymes studied in seven carnivores species, designation of loci and alleles, and alleles present in each species

Designa- Designa- Alleles present indicated with an asterisk

Enzyme tion of tionof
loci alleles Pine Beech
Redfox Stoat Weasel Polecat marten marten Badger

Acid phosphatase A CP 100 * *

85 * * * *
Adenosine deaminase Ada I50 * I *
100 t * * *
Adenylate kinase Ak 100 * * I * * *
Catalase Car 100 * *
50 * I *
Diaphorase Dia I 100 *
75 * * * * *
45 *
Dia I1 100 *
90 * * * *
63 *
Esterase Est I 150 * * * * *
100 * *
Est I1 140 * * * * *
127 *
100 *
Glucose-6-phosphate G6pd 107 * * *
dehydrogenase 100 * * *
73 *
Glutamic-oxaloacetic Got I loo * * * * * *
transaminase 88
Got I1 100 *
67 * *
20 * * *
Glutamic-pyruvic Gpt 100 * * * * *
transaminase 57 *
28 *
Isocitrate dehydro- led I 100 * * *
genase
Icd I1 100 * *
76 *
Lactate dehydrogenase Ldh 1 100 * *
100 *
Ldh I1 100 * * *
Malate dehydrogenase Mdh 1 I 70 * *
100 *
Mdh I1 118
100 * * *
Peptidase pep 100 *
78 * *
6-phosphogluconate Pgd 130 *
deh ydrogenase 110 *
100 *
Phosphoglycerate Pgk 133 *
kinase I10
100 * *
304 V . SIMONSEN Hereditas 96 (1982)

Table 5. The genetic identity (I) and distance (D) for the seven carnivore species

D for I for

Red Pole Pine Beech

fox Stoat Weasel cat marten marten Badger

Red fox 0.333 0.333 0.381 0.381 0.381 0.429

Stoat 1.099 0.762 0.810 0.619 0.619 0.333
Weasel 1.099 0.272 0.810 0.810 0.810 0.333
Pole cat 0.%5 0.211 0.21 I 0.714 0.714 0.429
Pine I .Ooo 0.333
0.%5 0.480 0.211 0.336
marten
Beech 0.336 0.000 0.333
0.965 0.480 0.211
marten
Badger 0.847 1.099 1.099 0.847 1.099 1.099

Acknowledgments. - I wish to thank Dr. 9 . Jensen for arrang- JENSEN. 9. (Ed.). 1976. Dansk vildtforskning 1975-76, p. 17
ing the contact with Mr. 0. K. Larsen, who has provided all the LEDOUX, R. G. and KENYON, A. J. 1975. Protides of the
samples used; Mrs. Doth Andersen and Mrs. Kirsten Petersen mustelidae. - 11. Immunologic relatedness. - Comp. B ~ o -
for excellent technical assistance; Mr. Terry Dobson for correct- chem. Physiol. 5 1 : 213-217
ing the manuscript; Mrs. Marianne Szygenda and Mrs. Lis Jen- MANDAHL, N . and FREDGA. K. 1980. A comparative chromo-
sen for typing the manuscript; and Mr. Arno Jensen for the some study by means of G-. C- and NOR-bandings of the
figure. weasel, the pygmy weasel, and the stoat, (Mustela, Carnivora,
Mammalia). -Hereditns 93: 7 5 4 3
MCDERMID, E. M., ANANTHAKRISHAN, R. and AGAR, N. S .
1972. Electrophoretic investigation of plasma and red cell pro-
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