ISSN 00329452, Journal of Ichthyology, 2012, Vol. 52, No. 11, pp. 821–837. © Pleiades Publishing, Ltd., 2012.

Morphological Diversity and Age Dependent Transformations

in Progeny of the Large Barbs (Barbus intermedius complex sensu
Banister) of Several Morphotypes from Lake Tana (Ethiopia).
Results of a LongTime Experiment1
M. V. Minaa, F. N. Shkila, K. F. Dzerzhinskiib, Belay Abdissac,
A. N. Mironovskyb, and D. V. Kapitanovaa
a
Koltsov Institute of Developmental Biology, Russian Academy of Sciences,
ul. Vavilova 26, Moscow, 119334 Russia
b Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences,
Leninskii pr. 33, Moscow, 119071 Russia
c Bahir Dar Fisheries and Other Aquatic Life Research Center, Bahir Dar, Ethiopia

email: mvmina@bk.ru
Received February 2, 2012

Abstract—Results of a longtime (2005–2011) experiment in crossing and rearing the large African barbs
from Lake Tana (Ethiopia) are presented. One may conclude that progeny from homonomic crossings of
largemouthed barbs acquire diagnostic traits of their parental morphotype at different age. Some males orig
inated from different crossings reached sexual maturity before acquiring diagnostic traits of a certain morpho
type. Siblings from both homonomic and heteronomic crossings living in the same conditions considerably
differed both in external and in skull characters. Differences both in external and skull characters between
some siblings originated from a homonomic crossing that lived in a pond and those that lived in aquaria were
equivalent to the differences between moiphotypes.
DOI: 10.1134/S0032945212110070
Keywords: species flock, morphotype, morphological diversity, reproductive isolation, ontogenetic channels

1
Large barbs inhabiting Lake Tana are extremely
morphologically diverse. As Nagelkerke et al. (1994)
wrote “The encountered morphological diversity par
allels that found in European cyprinids (ranging from
the deepbodied … bream, Abramis brama, to the spin
dleshaped … asp, Aspius aspius)” (p. 17). Differences
in external features provided the basis for describing a
number of species of the Lake Tana large barbs (Rüp
pell, 1836; Boulenger, 1902, 1911; Bini, 1940).
Nagelkerke et al. (1994) distinguished 13 morpho
types of the Lake Tana barbs and gave them names in
English. Later, Nagelkerke and Sibbing (2000) have
recognized 15 morphotypes as different species based
on data evidencing, in their view, that these forms are
reproductively isolated from each other. This decision
is arguable, and below we prefer to use a neutral term
“morphotype”.
It should be stressed that description of the mor
photypes/species was initially based on differences in
external features (coloration, shape of the body and
head, size and position of the mouth etc.) observed in
large individuals. Mina et al. (1993, 1996a) main
1 Article is published in the original.
tained that diagnostic traits of the moiphotypes were
acquired when fish were 3–5 years old at the standard
length (SL) 20–25 cm. At the same time, Nagelkerke
et al. (1995), though admitting that differentiation of
the morphotypes was the more distinct the larger the
fish, claimed that 11 of 14 distinguished morphotypes
could be identified at fork length <12 cm.
De Graaf et al. (2010) discussing causes of differ
ences between the morphotypes/species considered
two alternatives: (1) the differences are genetically
determined, (2) the differences are environmentally
2
induced . These authors carried out a “common gar
den experiment”, rearing progeny from homonomic
crosses of the morphotypes intermedius, troutlike and
bigmoth smalleye (which they recognize as species
Labeobarbus tsanensis, L. truttiformes and L. mega
stoma, respectively) in aquaria under the same condi
tions during two years. They found groups of offspring
of different origin to differ in external characters
(measurements) at SL < 40 mm and this, in their view,
2 This
view point for no apparent reason is ascribed to Mina et al.
(1996a, b, 2001)

821
822 MINA et al.

Table 1. Performed crossings

Crossing number Crossing Fertilization date
1 Troutlike × Troutlike 01/10/2005
2 Bigmouth smalleye × Bigmouth smalleye 01/10/2005
3 Bigmouth smalleye × Bigmouth smalleye 02/10/2005
4 Bigmouth smalleye × Bigmouth smalleye 03/10/2005
5 Intermedius × Intermedius 05/10/2005
6 Bigmouth bigeye × Bigmouth bigeye 06/10/2005
7 Intermedius × Intermedius 06/10/2005
8 Intermedius × Bigmouth bigeye 06/10/2005
9 Bigmouth bigeye × Acute 19/10/2005
10 Bigmouth bigeye × White hunch 19/10/2005
11 Intermedius × Intermedius 20/10/2007
12 Bigmouth smalleye × Intermedius 16/09/2008
13 Intermedius × (Bigmouth bigeye × Intermedius) 19/10/2007

favors the first alternative and is an evidence for the
biological species status of the morphotypes. In their
paper there are pictures of large (SL > 20 cm) individ
uals of the morphotypes under study, but not of the
progeny, and they did not compare progeny with large
individuals of the parental morphotypes/species.
In the present paper we discuss results of an exper
iment that lasted more than five years (from 2005 until
2011). In the course of the experiment we obtained
and reared progeny from crossings of barbs of the same
morphotype and of different morphotypes. Thus, we
could estimate morphological differences between
siblings and between offspring from different homo
and heteronomic crossings, as well as follow age
dependent changes of the morphological characters.
Special attention was paid to the characters that are
used for identification of the parental morphotypes.
MATERIALS AND METHODS
Progeny of the first generation was obtained cross
ing barbs caught during the spawning season in the
Gumara River flowing into Lake Tana and its tributary
the Ducalit River. For identification of morphotypes
we used descriptions and pictures presented by
Nagelkerke et al. (1994) and Nagelkerke and Sibbing
(2000). According to these authors morphotypes rep
resented in our experiment are characterized as fol
lows:
Acute—head longer than body depth, tapering and
narrow, terminal mouth, shallow body, silvery white,
sometimes with a slight nuchal hump (in larger speci
mens); the head with a straight to concave dorsal pro
file.
Bigmouth bigeye—very large eye (diameter equal
to snout length); lower jaw directed obliquely upward,
often extending beyond the upper jaw; terminal
mouth; the head with a slightly convex or concave dor
sal profile; nuchal hump.
Bigmouth smalleye—lower jaw directed upward,
extending beyond the upper jaw, nuchal hump; slightly
concave dorsal head profile; body depth less than head
length.
Intermedius—no extreme characters; inferior
mouth, the head with a slightly convex to slightly con
cave dorsal profile; the lower jaw shorter than the
upper jaw, when the mouth is closed.
Troutlike—spindleshaped, but relatively deep
body; head shorter than body depth, straight to con
cave dorsal profile; terminal mouth, the lower jaw is at
least as long as the upper jaw.
White hunch—nuchal hump (variable in size)
already present in very small specimens; body very
wide in pectoral area; wide head with straight to con
cave dorsal profile; lower jaw directed obliquely
upward; jaws equal in length; gape (sub)terminal.
Designations of the morphotypes in the following
text: Inintermedius (including the nominal species
B. intermedius Rüppell, B. tsanensis Nagelkerke &
Sibbing, B. brevicephalus Nagelkerke & Sibbing),
Be—bigmouth bigeye (described as Barbus mac
rophtalmus Bini), Se—bigmouth smalleye (B. mega
stoma Nagelkerke & Sibbing), Tr—troutlike (B. trutti
formis Nagelkerke & Sibbing), Wh—white hunch
(B. platydorsus Nagelkerke & Sibbing), Ac—acute
(B. acutirostris Bini).
Some fish caught during the spawning season in the
Gumara River were difficult to identify, and we
selected for crossings those individuals which could be
reliably affiliated with a certain morphotype.
Progeny reared at least for two years was obtained
from eight homonomic and five heteronomic crossings
(Table 1). In what follows, heteronomic crossings are

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS
labeled as (morphotype of the female) × (morphotype
of the male).
To fertilize eggs we placed them in the center of a
dry Petri dish, a small amount of milt (about l mL) of
one male was put close to the eggs but out of contact
with them. Then the water was poured into the dish,
and eggs and milt were carefully mixed. Several min
utes later, we washed out leftover milt and placed the
Petri dish in a trough with circulating aerated water or
at a shallow place in the river. Then fertilized eggs were
transferred to the indoor aquaria (20 L) with perma
nent aeration. The water temperature in the aquaria
was subjected to natural fluctuations inside the room
(17.6–19.0°C at 6–7 a.m. to 22.5–24.6°C at 2–
4 p.m.), the light regime was natural (12 hours light
and 12 hours dark). The aquaria bottoms were cleaned
every day changing 1/3 of water. The water was taken
from Lake Tana, treated with UV lamp and filtered
during 24 hours. Fish larvae were fed with live nauplii
of Artemia and artificial food Biomar (grains 0.2 mm).
When juveniles were 1–3 month old, they were trans
ferred to 180 L aquaria with the permanent aeration
and water circulation. The aquaria were located in a
room without heating, so the water temperature varied
in a wide range (20–26°C). Density of fish in aquaria
decreased in the course of experiment since some fish
died and some displaying obvious morphological
deformations were removed.
In May 2008, the fish were photographed and
tagged with microchips Tracer animal coder produced
by Bayer. The microchips were implanted under skin
near the dorsal fin using solution of benzocaine
(2 mL/5 L) as an anesthetic and Contraik (Tetra) for
disinfection before and after the implantation. After
tagging, the fish were placed for three days in an aquar
ium where Contraik and Stress Coat (Api) were added
to the water in concentrations used for prophylactic of
diseases. Later, when the fish were photographed, the
microchips were read using the scanner MiniMax II.
To photograph a fish we placed it in a separate 5 L
aquarium with movable rear wall and a visible rule. Pre
liminary treatment of the photographs and measure
ments of fish were performed using Photoshop CS2.
Some fish were transferred at different age to the
pond (5 m × 8 m, maximal depth 2 m) filled with water
from the lake. During the dry season the water level in
the pond was sustained by pumping lake water into it.
Larvae and juveniles of fish (Garra, Barbus), aquatic
invertebrates and plants arrived at the pond with the
lake water that was not treated or filtered. During the
wet season, the pond was filled with the rain water. The
fish were fed with Biomar and Tetra AniMin once in a
day. Fish from the pond were caught every six months
using a seine, photographed, then placed in quaran
tine for 2–3 days and returned to the pond. Some fish
were taken for preparation of skulls and estimation of
maturity stage according to Nikolsky (1963).
Eight measurements of each fish were made from
its photograph: SL—standard length (from the tip of
JOURNAL OF ICHTHYOLOGY Vol. 52 No. 11 2012
823
the snout to the base of the caudal fin), c—head
length, ao—snout length, lpc—caudal peduncle
length, o—eye diameter, H—maximal body depth,
cH—maximal head depth, h—caudal peduncle
depth. To minimize the measurement error, each mea
surement was repeated three times and the mean of the
three values was accepted as the measurement esti
mate.
Measurements of the skull and separate bones were
made according to the scheme presented in Fig. 1.
Shape of a bone was characterized by ratio (index) of
measurements of the bone. Characters of each cate
gory (external measurements, measurements of the
skull, measurements of separate bones) were made by
one and the same operator.
Indices characterizing shape of bones in reared
progeny were compared with those calculated for
skulls of large adult barbs collected by participants of
the Joint EthioRussian Biological Expedition in
1992–1994.
The principal component analysis (PCA) was per
formed on logtransformed bone shape indices as well
as on logtransformed absolute estimates of external
measurements and measurements of the skull exclud
ing SL or BL. Scatters were plotted in a system of
coordinates where the abscissa was SL (or BL), and
the ordinate—a principal component (PC1 or PC2).
Outlines of a scatter formed by progeny of a certain
group were regarded as boundaries of a fragment of an
“ontogenetic channel”, that is of an area in the char
acter space within which individual ontogenetic tra
jectories are located (Mina et al., 1996b; Mina, 2001).
Dealing with BL we calculated SL from the equa
tion SL mm = 4.107 BL mm + 29.306 mm (Mina et al.,
1996a).
PCA was carried out using NTSYS pc2.02k,
eigenvectors computed for correlation matrices,
length of vector was set equal to 1.
RESULTS
At the age 1+, in some broods there were males
with running milt (Table 2), most numerous in the
progeny In × In. In progeny from heteronomic cross
ings we found one running male Be × Wh and three Be ×
In. The milt of the latter ones was put into refrigerator
(t = +5°C) and next day used to fertilize eggs of an In
female caught in the Gumara River. The eggs were
divided in three batches, fertilized with the milt of dif
ferent males, and incubated under standard condi
tions. Mortality of the eggs and larvae at early stages of
development did not differ from that observed in prog
eny from homonomic crossings. Survived larvae were
reared and then transferred to the pond where the fish
lived till 2010. Among seven fish taken from the pond
in 2010 there were two running males and two females
with gonads at maturity stages III and IV.
824 MINA et al.

(a) (b)
(c)
Po
B1
l
B3 BL
B4 HS1
2
HS2
B2
(d) l

2, Op
l (f) l
(e)

2, Hm
2, Io

(h) l
l
(g)
l* 2* (i)
l

2, Mx 2, Pmx 2, De
Fig. 1. Scheme of measurements of the skull and individual bones in large African barbs (Barbus intermedius complex); shape of
a bone is characterized by ratio of a measurement 1 to measurement 2; (a) skull dorsal view: B1, B2, B3—distances between outer
margins of frontals, pterotics, and sphenotics, respectively; B4—skull width at the level of junction of frontals and pterotics; (b)
skull lateral view: BL—basal length of the skull, HS1, HS2—skull depths at the level of the bend of parasphenoid and posterior
margin of parasphenoid, respectively; (c) preopercle (1—depth of vertical ramus, 2—length of horizontal ramus, 3—diagonal
length); (d) opercle (1—diagonal distance between anterior superior angle and posterior inferior angle, 2—depth along the ante
rior margin of the bone); (e) interopercle (1—maximal depth, 2—maximal length); (f) hyomandibular (1—length of the dorsal
junction, 2—maximal depth); (g) dentary (1—maximal depth, 2—maximal length); (h) maxilla (1—maximal depth, 1*—min
imal depth behind the palatine process, 2—maximal length, 2*—minimal depth in front of the palatine process); (i) premaxilla
(1—depth of the process, 2—maximal length of the bone).

External Appearance
We see no sense to estimate frequency of morpho
logical variants in progeny from experimental cross
ings. On the one hand, taken into account small num
bers of the progeny, it seems probable that observed
morphological diversity was much less than that in the
nature, and on the other hand, some morphological
variants observed in the experiment might emerge in
the experiment, but do not emerge in the nature.
Thus, all we can do is to state that emergence of a cer
tain morphological variant in progeny from a given
crossing is possible.
Homonomic Crossings
Bigmouth bigeye × Bigmouth bigeye (crossing
no. 6). Fish lived in an aquarium. All of them regard
less of their age had eyes bigger than progeny of the
same size originated from other crossings. Other diag
nostic traits of the parental morphotype were not
acquired until the fish were three years old (Fig. 2a). At
the age 5+ all three survived individuals could be iden
tified with the parental morphotype (Fig. 2b).
Bigmouth smalleye × Bigmouth smalleye (cross
ings nos. 2, 3, 4). At the age 5+ some fish that lived in
aquaria displayed the diagnostic traits of the parental
morphotype (Fig. 2c), whereas their siblings did not
(Fig. 2d).Ten individuals lived in the pond from 2007
till 2009, and they differed drastically both from their
siblings that lived in aquaria and some of them from
their parents in having elongated snout and quite dis
tinct nuchal hump (Fig. 2e).

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS 825

Table 2. Proportion of running males in broods at the age of 1+

Number of running males/size SL of running
Crossing Fertilization date
of the brood males, mm
1. Troutlike × Troutlike 01/10/2005 0/7 –
2. Bigmouth smalleye × Bigmouth smalleye 01/10/2005 1/16 103
3. Bigmouth smalleye × Bigmouth smalleye 02/10/2005 1/5 96
4. Bigmouth smalleye × Bigmouth smalleye 03/10/2005 1/7 102
5. Intermedius × Intermedius 05/10/2005 12/21 95*
6. Bigmouth bigeye × Bigmouth bigeye 06/10/2005 1/22 125
7. Intermedius × Intermedius 06/10/2005 4/11 83*
8. Bigmouth bigeye × Intermedius 06/10/2005 3/10 112*
9. Bigmouth bigeye × Acute 19/10/2005 0/5 –
10. Bigmouth bigeye × White hunch 19/10/2005 1/12 104
* mean SL.

Troutlike × Troutlike (crossing no. 1). Progeny of
this crossing displayed no obvious traits of the parental
morphotype (Fig. 2f).
Intermedius × Intermedius (crossings nos. 5, 7, 11).
The diagnosis of the morphotype Intermedius is rather
vague. The main characteristic of this morphotype is
the absence of “extreme characters” that is diagnostic
characters of other morphotypes. Accepting the given
diagnosis we could identify all the progeny from the
crossings In × In with the parental moiphotype. Nev
ertheless, differences between individuals originated
from the crossings were obvious: some of them had
blunt snout and the mouth subinferior (Fig. 3a), in
others the snout was pointed and the mouth terminal
(Fig. 3b), or subsuperior (Fig. 3c), the nuchal hump was
absent (Figs. 3a, 3b), or present (though slight) (Fig. 3c).
Heteronomic Crossings
Bigmouth smalleye × Intermedius (crossing
no. 12). Fish lived in an aquarium. At the age 1+ they
did not display characters of the morphotype Se being
rather similar with some representatives of the mor
photype In (cf. Fig. 3d and Fig. 3c).
Bigmouth bigeye × White hunch (crossing no. 10).
Fish lived in an aquarium. Among the progeny from
this crossing no individuals without obvious morpho
logical deformations reached the age 5+. At the age 3+
and 4+ the fish were rather similar with the Be × Be
progeny of the same age and size (cf. Fig. 3e and Fig. 2a).
There was nothing in their appearance to suggest that
their father was Wh.
Bigmouth bigeye × Acute (crossing no. 9). Fish
lived for three years in an aquarium and then trans
ferred to the pond. The characters of the parents were
bizarrely combined in them, and they could not be affili
ated with any of the described morphotypes (Fig. 3f).
Bigmouth bigeye × Iintermedius (crossing no. 8).
Fish lived in an aquarium. One individual acquired
specific traits of the morphotype Be at the age of two
years (Figs. 4a, 4b), whereas its siblings had no such
traits even at the age 5+ (Fig. 4c).
Intermedius × (Bigmouth bigeye × Intermedius)
(crossing no. 13). Fish lived in an aquarium until the
age 1+ and then transferred to the pond. At the age 2+
they had no characters of the morphotype Be and
could be sooner identified with In (Fig. 4d).
External Measurements
In scatterplots considered below some individuals
are represented with several points because they were
measured repeatedly in the course of the experiment.
Individuals kept in aquaria during the whole life
form two ontogenetic channels in the system of coor
dinates SL and the PC2 (Fig. 5). Within the lower
channel we find the progeny from the crossings In ×
In, and within the upper one—the progeny from
crossings Be × Be, Se × Se, and Tr × Tr, morphotypes
assigned by Mina et al. (2001) to the “largemouthed”
phenetic group. The channels overlap until the fish
reach SL ≈ 14 cm, and are completely divided at SL >
14 cm (age ≥2+). One In × In offspring of SL ≈
229 mm (pictured in Fig. 2b) as an evident outlier was
not considered depicting the In × In channel. The PC2 is
loaded mainly by the characters ao, o, and H (Table 3).
The indices ao (i.e. ao/SL) and o (o/SL) are nega
tively correlated with SL. At the same SL, their esti
mates, as a rule, are the biggest in the progeny of Be ×
Be and the smallest in In × In, but in some fish of the
former group at SL > 18 cm these estimates are equal
to or smaller than in the fish of the latter group at SL <
14 cm.
Nagelkerke et al. (1994) and Nagelkerke and Sib
bing (2000) suggested recognizing the morphotype Be
by the value of the ratio o/ao approximately equal to or
greater than 1 in Be and smaller than 1 in other large
mouthed barbs. Yet, according to our data, this index

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826 MINA et al.

(d)
(b)

(f)
(a)

(e)
(c)

Fig. 2. (a) An offspring from the crossing Be × Be at the age 3+, (b) the same individual at the age 5+, (c, d) offspring from the
crossing Se × Se that lived in aquaria at the age 5+; (e) their sibling from the pond at the age 3+; (f) an offspring from the crossing
Tr × Tr at the age 5+.

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS 827

(d)
(b)

(f)
(e)
(c)
(a)

Fig. 3. (a, b, c) Offspring from the crossings In × In ((a, b) at the age 5+, (c) at the age 3+); (d) an offspring from the crossing Se × In at
the age 1+; (e) an offspring from the crossing Be × Wh at the age 1+; (f) an offspring from the crossing Be × Ac at the age 4+.

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828 MINA et al.

(d)
(b)

(c)
(a)

Fig. 4. (a) An offspring from the crossing Be × In at the age 1 year, (b) same fish at the age 2 years, (c) its sibling at the age 4+,
(d) an offspring from the crossing In × (Be × In) at the age 2+.

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS 829

PC2
–0.22

–0.30

() I
() B
–0.37 () S
() T

–0.45

–0.53
81.60 130.95 180.30 229.65 279.00
SL, mm

Fig. 5. External measurements of progeny from the crossings In × In (I), Be × Be (B), Se × Se (S), and Tr × Tr (T). Scatterplot of
the PC2 against SL.

is also negatively correlated with SL, and it is possible pond, so it is not quite correct to compare them with
to distinguish Be from Se and Tr only considering fish the fish from aquaria.
of the same size (Fig. 6). Scatters of Se × Se progeny that lived in aquaria and
In scatterplots of PC2 against SL, the progeny from those which lived for two years in the pond overlap in
the crossings Be × Be, Se × Se, and Tr × Tr, both taken the PC2 (Fig. 8) the most of fish from the pond having
together and by pairs, did not form separated ontoge smaller o and bigger lpc than their siblings from
netic channels. aquaria (Table 3). The fish in the pond grew faster than
in aquaria, so most of the former ones being four years
We can follow changes in the external characters of old were bigger than their siblings from the aquaria at
two fish originated from the crossing Be × In (crossing the age of 5+.
no. 8, Table 1). They were measured in October 2006,
August 2007, February 2010, and one of them also in
March 2011. Thus, in Fig. 7 one offspring is repre Measurements of the Skull
sented with three points and another with four. Con
necting the points representing the same individual we Just as in the case of external measurements, two
obtained the individual ontogenetic trajectory. These ontogenetic channels are separated in the scatterplot
trajectories, as well as three points representing back
crosses In × (Be × In) measured in October 2007, were Table 3. Eigenvector loadings of external measurements on
plotted along with the points representing the progeny the PC2
from crossings Be × Be and In × In that formed onto
genetic channels diverging at SL ≈ 14 cm. These chan Scatterplot
nels are divided mainly due to loadings of the mea Character
surements o and ao (Table 3). At the age of one year the Fig. 5 Fig. 7 Fig. 8
hybrids Be × In fall in the In × In channel and later c 0.2405 0.1991 0.0855
take up an intermediate position between the Be × Be
ao 0.5520 0.4281 –0.1005
and In × In channels, whereas backcrosses occur
within the channel Be × Be, Analysis of individual lpc –0.2308 –0.3318 –0.5805
characters shows that the hybrids are closer to Be in o 0.4697 0.6107 0.7962
ao, and closer to In in o. H –0.4462 –0.4205 0.0011
The only offspring from the crossing Se × In mea cH –0.2312 –0.1481 –0.0340
sured in October 2007 falls within the Se × Se channel. h –0.3331 –0.3087 –0.1025
Yet, it should be noticed that this fish, as well as the
backcrosses In × (Be × In), spent part of their life in the Note: Designations of characters are given in Material and methods.

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830 MINA et al.

o/ao
126

1.13
() B
() S
() T
1.01

0.89

0.77
82.40 121.02 159.64 198.26 236.88
SL, mm

Fig. 6. The relationship between the ratio o/ao and SL in progeny from the crossings Be × Be (B), Se × Se (S), and Tr × Tr (T).

PC2
–0.22

() I
–0.30 () B
() BC
() Ha
–0.37 () Hb

–0.45

–0.53
81.60 130.95 180.30 229.65 279.00
SL, mm

Fig. 7. External measurements of progeny from the crossings In × In (I), Be × Be (B), Be × In (Ha and Hb), and In × (Be × In)
(BC). Scatterplot of the PC2 against SL. Individual trajectories of Ha and Hb are traced.

PC2 and SL before progeny from homonomic cross
ings reach BL ≈ 2.5 cm (SL ≈ 13 cm), the offspring
In × In falling into the upper channel, and the remain
ing ones—into the lower one (Fig. 9).
The main loadings on the PC2 are from the charac
ters B1, De, and Io (Table 4). The progeny from cross
ings Be × Be, Se × Se, and Tr × Tr taken together and
by pairs do not form separated channels.
The progeny Be × In and backcrosses In × (Be × In)
are found between the Be × Be and In × In channels
(Fig. 10), the PC2 being loaded mainly by B1, De, and
Io (Table 4).

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS 831

PC2
–0.12

() s4
() s5
–0.16
() s6
() sp

–0.21

–0.26

–0.30
134.20 168.18 202.16 236.15 270.13
SL, mm

Fig. 8. External measurements of progeny from the crossing Se × Se that lived in aquaria during four (s4), five (s5) and six (s6)
years and those that lived during one year in the aquarium and during two years in the pond (sp). Scatterplot of the PC2 against SL.

PC2
0.07

0.01

–0.05

() I
–0.12 () B
() S
() T

–0.18
17.10 26.30 35.55 44.75 54.00
BL, mm

Fig. 9. Skull measurements of progeny from the crossings In × In (I), Be × Be (B), Se × Se (S), and Tr × Tr (T). Scatterplot of the
PC2 against BL.

The scatters of the progeny from the crossings Se × Shape of Bones

Se that lived in aquaria and of those which spent two
years in the pond overlap, the latter being shifted to the Division of ontogenetic channels In × In and Be ×
area of smaller PC2 scores (Fig. 11) mainly due to Be + Se × Se is evident in the scatterplot of the PC1
loadings of B1, Io, and HS1 (Table 4). against BL (Fig. 12). Divergence of the channels takes

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832 MINA et al.

Table 4. Eigenvector loadings of skull measurements on the DISCUSSION

PC2
Earlier it was shown that at the age 3+ and SL =
Scatterplot 12–18 cm some offspring from the crossings Be × Be,
Character
Fig. 9 Fig. 10 Fig. 11 Se × Se, and Tr × Tr displayed no external features of
their parental morphotypes (Mina et al., 2010). Now,
B3 0.0862 0.0096 0.0333 the same can be stated regarding the progeny Se × Se
B2 0.0984 0.1178 –0.0587 and Tr × Tr at the age 5+ and SL = 20–22 cm. Some
B1 0.6845 0.7197 –0.6527 individuals could not be classified with a certain mor
photype, but could be assigned to the “large
B4 0.2501 0.2024 –0.2092 mouthed” phenetic group, while others, if they were
HS1 0.2286 0.2273 –0.4479 caught in the nature, would be, most probably, identi
HS2 0.1212 0.1065 –0.0237 fied as In—the morphotype without “extreme charac
Hm –0.0611 –0.0259 0.0652 ters”. Now, is it possible to predict to what morpho
type such fish would be assigned at an older age if they
Pm –0.2567 –0.2673 0.2189 lived longer?
Po –0.1089 –0.0829 0.1568
We may suppose that a fish, which during the
Op –0.0834 –0.0368 0.1567 experiment acquired some specific traits of the paren
Io –0.3448 –0.3636 0.4106 tal morphotype, would, probably, acquire with the age
Mx –0.1825 –0.1664 0.2318 its other traits and would be eventually classified with
De –0.3798 –0.3520 0.0283 this morphotype, but if a fish during the experiment
did not acquire specific traits of a certain morphotype,
Note: B1, B2, B3—distances between outer margins of frontals, we can not make any decision concerning its morpho
pterotics, and sphenotics, respectively; B4—skull width at the
level of junction of frontals and pterotics; HS1, HS2—skull type at an older age. May be, it would acquire charac
depths at the level of the bend of parasphenoid and posterior ters of the parental morphotype later, and may be it
margin of parasphenoid, respectively; Hm—depth of hyo would not.
mandibular, Pm—length of premaxilla, Po—length of preo
percle measured along the diagonal, Op—depth of the ante It is worth mentioning that whereas some individu
rior margin of opercle, Io—length of interopercle, Mx— als among the progeny Se × Se could be confidently
length of maxilla, De—length of dentary. affiliated with the morphotype Se (Fig. 2c), none of
the progeny Tr × Tr displayed distinct external traits of
the morphotype Tr and could be readily assigned to the
place at BL = 2.6 cm (SL ≈ 13 cm), the PC1 is loaded morphotype Se (Fig. 2f). This seems to be the reason
mainly by the indices iPm and iMx (Table 5). Besides, why Nagelkerke et al. (1995) could not find in the
the progeny Tr × Tr form a separate channel. nature Tr individuals at SL < 20 cm.
In the scatterplot where not only the progeny, but Our judgment on phenetic relationships of barbs
also adult individuals Tr, Be, and Se are represented, originated from different crossings greatly depends on
all the adults form a cluster separated from all the a set of characters we use. The PCA of the external and
progeny in the PC1, but in the PC2 the Tr × Tr channel skull measurements allowed us to distinguish the prog
is shifted in relation to the common channel of Be × eny In × In and largemouthed barbs at SL > 14 cm.
Be + Se × Se, and the Tr adults occur in the Tr × Tr The latter ones formed a joint scatter in the coordi
channel, whereas the adults Be and Se fall into the nates SL (or BL) and the PC2, but we could not reveal
channel Be × Be + Se × Se (Fig. 13). The PC2 in this separate ontogenetic channels of Be × Be, Se × Se, and
case is loaded mainly by the indices iPm and iMx, that Tr × Tr analyzing them either taken together or by
is by the same indices that load the PC1 in Fig. 12 pairs. The PCA of indices characterizing shape of
(Table 5). bones also showed that in the coordinates BL and the
PC1 the scatter of the In × In progeny was separated
All three studied hybrids Be × In and two of from that of the largemouthed barbs, though not so
10 backcrosses fall into the channel of In × In, whereas distinctly as in the case of external and skull measure
eight backcrosses are closer to the Be × Be channel ments. Besides, the scatter of Tr × Tr occurred to be
(Fig. 14), the PC1 is loaded mainly by iPo and iHm nearly completely separated from that of Be × Be + Se ×
(Table 5). Se, mainly due to differences in the indices iMx and
iPm which were smaller in Tr × Tr. Since the similar
The progeny from the crossings Se × Se that lived differences are found between large adult individuals
for two years in the pond distinctly differ in the PC1 of the morphotypes Be + Se and Tr (Fig. 13), we sug
scores from their siblings of similar size (BL > 3.7 cm, gest that if progeny from experimental crossing Tr × Tr
SL > 18 cm) which lived in aquaria (Fig. 15). Main lived longer they would, probably, acquire other spe
loadings on the PC1 are from the indices iIo, iDe, and cific traits of their parental morphotype, at least, they
iPm (Table 5). would not be affiliated with the morphotypes Be or Se.

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS 833

PC2
0.12

0.07

0.05 () I
() B
() BC
() HY
–0.02

–0.06
25.80 32.85 39.90 46.95 54.00
BL, mm

Fig. 10. Skull measurements of progeny from the crossings In × In (I), Be × Be (B), Be × In (HY), and In × (Be × In) (BC). Scat
terplot of the PC2 against BL.

PC2
0.13

( ) s4
0.09
( ) s5
( ) s6
( ) sp
0.05

0.01

–0.03
26.70 33.55 40.45 47.30 54.20
BL, mm

Fig. 11. Skull measurements of progeny from the crossing Se × Se that lived in aquaria during four (s4), five (s5) and six (s6) years
and those that lived during one year in the aquarium and during two years in the pond (sp). Scatterplot of the PC2 against SL.

Those males originated from both homonomic and
heteronomic crossings which displayed no specific
traits of their parental morphotype(s), but were ripe
and running would be hardly recognizable among run
ning males of SL ≤ 15 cm numerous at the spawning
grounds in the Gumara River and its tributaries. Such
males may enter into the composition of “unit groups”
of spawning fish together with larger males and
females of different morphotypes (Dzerzhinskii et al.,
2007) and are identified as “precocious intermedius”

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834 MINA et al.

PC1
2.89

2.84

2.78

() I
() B
2.73 () S
() T

2.67
17.10 26.30 33.55 44.75 54.00
BL, mm

Fig. 12. Indices characterizing shape of bones in progeny from the crossings In × In (I), Be × Be (B), Se × Se (S), and Tr × Tr (T).
Scatterplot of the PC1 against BL.

(Alekseyev et al., 1996) or Barbus brevicephalus sensu Up to now, there are no data allowing reliable eval
Nagelkerke and Sibbing (2000) based mainly on their uation of the degree of reproductive isolation between
size and ripeness. barbs assigned to different morphotypes. It was shown
that, at least between some morphotypes, there was no
strict premating isolation (Dzerzhinskii et al., 2007).
Table 5. Eigenvector loadings of bone shape indices on the As for the postmating isolation, mortality of progeny
PC1 or PC2 from different homonomic and heteronomic crossings
on early stages of development was, more or less, the
Scatterplot same. At the same time, mortality of the progeny from
Character Fig. 12 Fig. 13 Fig. 14 Fig. 15
crossings of females In and Se with males Varicorhinus
beso, a species closely related to barbs, at early stages
PC1 PC2 PC1 PC1
of development approached 100% (Dzerzhinskii et al.,
iPo 0.2542 0.2396 0.5423 0.3382 2007).
iOp –0.1826 0.3674 0.2059 0.2124 We can not estimate mortality of progeny from dif
ferent crossings before and after sexual maturation, as
iIo 0.3507 –0.0234 0.4881 0.4792
well as their fertility, but it is noteworthy that males
iHm 0.3545 –0.0383 0.4373 0.3552 originated from heteronomic crossings Be × Wh and
iDe 0.2982 0.0132 0.4031 –0.3846 Be × In reached the sexual maturity, and that crossing
iMx 0.5071 –0.5505 0.2225 0.2558 the latter with an In female caught in the Gumara
River we obtained viable progeny that also reached
iMxh –0.0588 0.3716 0.1353 0.3439
sexual maturity.
iPm 0.5512 –0.6035 –0.0611 0.3910 Some progeny from heteronomic crossings, e.g.
Note: iPo—ratio of the depth of the vertical ramus of preoprcle those originated from the crossing Be × Ac, looked
to the length of its horizontal ramus, iOp—ratio of the rather strange (Fig. 3f), but we saw not less strange
diagonal distance between anterior superior angle and pos looking barbs caught in the nature and they seemed to
terior inferior angle of opercle to the depth along the ante
rior margin of the bone, iIo—ratio of the maximal depth of be quite viable.
interopercle to its maximal length, iHm—ratio of the Judging by the studied progeny from the hetero
length of the dorsal junction of hyomandibular to the max nomic crosses Be × In, Se × In, and Be × Wh, hybrids
imal depth of the bone, iDe—ratio of the maximal depth of morphotypes can hardly be identified in the nature.
of dentary to its maximal length, iMx—ratio of the maxi
mal depth of maxilla to its maximal length, iMxh—ratio of While hybrids Be × Wh and some Be × In (Figs. 3e, 4b)
the minimal depth of maxilla behind the palatine process had appearance allowing to suppose that there were Be
to the miinimal depth in front of it, iPm—ratio of the pro individual(s) among their ancestors, some Be × In
cess of premaxilla to the maximal length of the bone. (Fig. 4c) and backcrosses In × (Be × In) (Fig. 4d) being

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 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS 835

PC2
0.06 () B
() S
() T
() Be
–0.01 () Se
() Tr

–0.08

–0.15

–0.22
17.10 32.35 47.65 62.90 72.80
BL, mm

Fig. 13. Indices characterizing shape of bones in progeny from the crossings Be × Be (B), Se × Se (S), Tr × Tr (T), and in large
individuals of the parental morphotypes (Be, Se, Tr). Scatterplot of the PC2 against BL.

PC1
3.53

3.48

3.42

() I
3.37 () B
() BC
() HY

3.32
25.50 32.60 39.75 46.85 54.00
BL, mm

Fig. 14. Indices characterizing shape of bones in progeny from the crossings In × In (I), Be × Be (B) Be × In (HY), and In × (Be ×
In) (BC). Scatterplot of the PC1 against BL.

caught in the nature would be most probably affiliated
with the morphotype In.
Analysis of external and skull measurements as well
as of indices characterizing shape of bones may help
revealing hybrid origin of barbs, but the validity of this
approach can be evaluated only after studying much
bigger material than that in our disposal.
The question: “are differences between the mor
photypes of the Lake Tana barbs genetically deter
mined or environmentally induced?”, in our view,
should be considered with regard to concrete morpho
types, since there is no reason to believe that all the
differences between morphotypes are of the same
nature. We also believe that it is a question of degree,

JOURNAL OF ICHTHYOLOGY Vol. 52 No. 11 2012

836
PC1
2.88
2.84
2.79
2.75
2.70
26.70 33.55
Fig. 15. Indices characterizing shape of bones in progeny from the crossing Se × Se that lived in aquaria during four (s4), five (s5)
and six (s6) years and those that lived during one year in the aquarium and during two years in the pond (sp). Scatterplot of the
PC1 against BL.
not “either–or”. Our data indicate that the diagnostic
characters of the described morphotypes may strongly
depend on environmental conditions. For instance,
differences in appearance and in indices characteriz
ing bones’ shape between siblings originated from the
crossings Se × Se that lived during four years in aquar
ium and those that lived for two years in the pond are
equivalent to differences between morphotypes. At the
same time, we could see that siblings living at the same
environmental conditions substantially differed in
their morphology. In this connection, we can recollect
that the siblings In × In were shown to differ in their
reaction toward change in the level of the thyroid hor
mone: some of them displayed obvious morphological
transformations and some did not (Shkil et al., 2010).
Such differences between siblings are usually consid
ered as genetically determined, but it is quite plausible
that their degree depends on the environmental condi
tions, and in the nature the differences may be greater
or smaller than in the experiment.
CONCLUSIONS
Though many papers are dedicated to the Lake
Tana barbs, the ways and mechanisms of their evolu
tion that has culminated in extraordinary phenotypic
diversity are still not clearly understood. We believe
that experimental crossing of barbs both of the same
and of different morphotypes and longtermed obser
vations of the progeny might greatly help to gain
insight into the situation. Ideally, barbs of all the mor
MINA et al.
() s4
() s5
() s6
() sp
40.45 47.30 54.20
BL, mm
photypes should be used in such studies, but this is
hardly possible due to technical difficulties.
The data presented above are scarce, and our infer
ences should be cautiously extrapolated to the natural
situation especially dealing with morphotypes that
were not represented in our experiment. Nevertheless,
we find it possible to state that:
(1) Progeny from homonomic crossings of large
mouthed barbs acquire diagnostic traits of their paren
tal morphotypes at different age, and some of them do
not acquire such traits even at the age 5+.
(2) Among progeny from both homonomic and
heteronomic crossings some males which do not dis
play the diagnostic traits of their parental morpho
type(s) reach sexual maturity and are fertile.
(3) Identification of offspring from heteronomic
crossings and backcrosses by external appearance is
difficult if not impossible.
(4) Some siblings living in different environmental
conditions substantially differ both in external and
skull characters.
(5) In some cases, the differences between siblings
both in external and skull characters may be equivalent
to the differences between recognized morphotypes.
ACKNOWLEDGMENTS
This study was performed within the framework of
the Joint EthioRussian Biological Expedition
(JERBE III) with the financial support from the Rus
sian Foundation for Basic Research (projects nos. 08
JOURNAL OF ICHTHYOLOGY Vol. 52 No. 11 2012
 MORPHOLOGICAL DIVERSITY AND AGE DEPENDENT TRANSFORMATIONS
0400061a, 100400787a, 110400117a, 1104
00109a) and the Program of the Presidium of the Rus
sian Academy of Sciences “Biodiversity and Dynam
ics of the Gene Pool”. We are grateful to S.V. Smirnov,
A.A. Darkov, D.V. Shchepotkin, Wondie Zelalem,
Mesfin Tsegaw, and the director and all the staff of the
BahirDar Fishery and Other Aquatic Life Research
Center (Ethiopia) for their help in the organization
and realization of this research and to Yu.Yu. Dge
buadze for valuable comments on the manuscript.
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