Journal of Fish Biology (1996) 48, 714–725

Environmental determination of sex in Apistogramma

(Cichlidae) and two other freshwater fishes (Teleostei)
U. R̈  W. B
University of Bielefeld, Department of Biology, Postfach 100131, 33501 Bielefeld,
Germany

(Received 15 July 1994, Accepted 30 June 1995)

Environmental sex determination by temperature could be revealed significantly in 33

Apistogramma-species and in Poecilia melanogaster. In some, but not all, Apistogramma-species
pH also influences the sex ratio, whereas neither temperature nor pH affect the sex ratio of
Pseudocrenilabrus multicolor victoriae. The sex in offspring of A. trifasciata is determined
within a sensitive period of about 30 to at least 40 days after spawning.
? 1996 The Fisheries Society of the British Isles

Key words: Apistogramma; Cichlidae; environmental sex determination; pH; sex ratio;
temperature.

INTRODUCTION
Sex is determined in reptiles by at least two different mechanisms, one genotypic
and the other environmental (Bull, 1980). In fish a genetic mechanism for sex
determination was proposed long ago by Aida (1921). Since then it has been
established for most species which have been examined (Price, 1984). Determi-
nation of sex by environmental factors after conception seems to be rare.
Environmental factors such as pH in some cichlids and a poecilid (Rubin, 1985),
and temperature in Menidia menidia (L.) (Conover & Kynard, 1981), M.
peninsula Goode & Bean (Middaugh & Hemmer, 1987) and Poeciliopsis lucida
Miller (Sullivan & Schultz, 1986; Schultz, 1993) have been shown to influence sex
ratio, too. Hints in the aquarium literature motivated our investigations of the
influence of temperature and pH on sex ratio in Apistogramma (Teleostei,
Cichlidae). The objective of this article is to present further data in regard to
environmental sex determination (ESD) in fish as this seems to be more common
than currently appreciated.

MATERIALS AND METHODS

Thirty-seven species of Apistogramma, a genus of neotropical cichlids, Pseudo-
crenilabrus multicolor victoriae Seegers, an African mouth-breeding cichlid, and Poecilia
(Limia) melanogaster (Günther), a Jamaican live-bearing poecilid (Table I), were used for
this study. Nomenclature of Apistogramma-species follows Ufermann et al. (1987). All
fish were of known geographical origin which might be of special importance for further
ecological investigation.
Generally, specimens taken from the wild and their F1-progeny were used for the
experiments with Apistogramma-species. There was no significant difference in sex ratio
of offspring in specimens from different imports or strains of long-time inbred laboratory-
stocks (F12 up to F15). Pseudocrenilabrus m. victoriae were F1-progeny of specimens
714
0022–1112/96/040714+12 $18.00/0 ? 1996 The Fisheries Society of the British Isles
 T I. Mean percentage of males within broods of 39 Teleostei

23) C 26) C 29) C

Species
pH 4·5 pH 5·5 pH 6·5 pH 4·5 pH 5·5 pH 6·5 pH 4·5 pH 5·5 pH 6·5

Apistogramma
agassizii (Steindachner) 42·2 (1) 77·3 (7) — 60·2 (1) — — 90·1 (1) 73·0 (6) —
borellii (Regan) 39·6 (1) 37·1 (3) 32·8 (1) 68·4 (2) 53·9 (2) 68·4 (2) 85·4 (1) 73·6 (4) 61·4 (1)
cacatuoides Hoedeman — 19·8 (5) — 84·3 (5) 62·7 (5) 43·3 (5) — 83·0 (5) —
caetei Kullander 59·9 (5) 53·3 (12) 3·8 (1) 43·7 (3) 48·0 (14) 14·5 (8) 37·5 (3) 52·0 (9) 4·4 (10)
diplotaenia Kullander 55·4 (1) 43·9 (3) — — 53·0 (2) — 98·8 (1) 79·9 (4) —
eunotus Kullander 44·7 (2) 38·5 (2) 31·1 (2) 63·4 (1) 53·4 (3) 41·2 (2) 76·6 (2) 64·1 (2) 66·8 (2)
geisleri Meinken — 28·7 (2) — — 50·9 (3) — — 82·4 (2) —
gephyra Kullander 45·8 (3) 40·6 (3) 35·5 (3) 65·7 (3) 56·1 (3) 39·2 (3) 78·0 (3) 62·4 (3) 66·8 (3)
gibbiceps Meinken — 19·2 (3) — 68·9 (2) 53·6 (2) 35·1 (2) 92·6 (2) 82·0 (2) 76·0 (2)
gossei Kullander — 28·7 (2) — — 49·2 (2) — — 81·4 (2) —
hippolytae Kullander 37·7 (1) 53·3 (2) — — 49·4 (1) — 93·2 (1) 85·7 (2) —
hoignei Meinken — 30·9 (3) — — — — — 81·7 (4) —
hongsloi Kullander 35·4 (3) 29·3 (3) 23·6 (3) 61·6 (3) 27·5 (3) 14·4 (3) 92·8 (3) 79·6 (3) 81·5 (4)
inconspicua Kullander 38·9 (2) 27·2 (2) 16·7 (2) — — — — 76·8 (2) —
iniridae Kullander — 34·4 (3) — — — — — 79·6 (2) —
linkei Koslowski — 15·9 (17) — 66·2 (7) 49·2 (7) 34·2 (6) — 88·9 (13) —
luelingi Kullander — 30·0 (2) — — — — — 88·5 (2) —
macmasteri Kullander — 30·4 (2) — — 50·9 (2) — — 86·7 (2) —
meinkeni Kullander 47·5 (1) 37·6 (2) 40·4 (1) 60·3 (1) 55·4 (2) 45·9 (1) 68·7 (1) 65·2 (2) 60·9 (1)
 T I. Continued

23) C 26) C 29) C

Species
pH 4·5 pH 5·5 pH 6·5 pH 4·5 pH 5·5 pH 6·5 pH 4·5 pH 5·5 pH 6·5

Apistogramma
mendezi Römer — 35·8 (2) — — — — — 87·2 (2) —
nijsseni Kullander 14·4 (14) 9·9 (12) 7·5 (13) 58·7 (20) 49·6 (24) 45·9 (21) 94·8 (12) 85·0 (18) 75·5 (16)
norberti Staeck 31·9 (2) 27·3 (2) 15·7 (2) 71·4 (2) 53·3 (2) 31·1 (2) — 89·2 (3) —
ortmanni (Eigenmann) — 25·0 (2) — — 48·8 (1) — — 82·9 (2) —
paucisquamis Kullander & Staeck 43·2 (2) — 31·1 (2) 67·2 (1) 52·9 (1) 41·1 (1) 76·3 (2) — 67·0 (2)
pertensis (Haseman) — 29·3 (6) — — — — — 76·4 (6) —
resticulosa Kullander 50·5 (2) — 25·7 (2) — — 38·7 (2) 90·3 (2) — 75·9 (2)
staecki Koslowski 27·8 (2) — — 54·3 (2) — — 91·5 (2) — —
steindachneri (Regan) 41·1 (3) — 19·3 (3) — — — 90·9 (4) — 77·0 (2)
trifasciata (Eigenmann & Kennedy) — 16·8 (15) — — 49·0 (15) — — 85·9 (15) —
uaupesi Kullander 34·9 (3) 26·4 (3) — 81·7 (3) 58·8 (2) — 97·5 (3) 83·8 (2) —
‘ Breitbinden ’ sp.* — 26·3 (5) — 79·2 (2) 68·5 (2) — — 89·3 (5) 65·0 (1)
‘ Gelbwangen ’ sp.* — 31·1 (3) — — — — — 70·6 (2) —
‘ Orangeschwanz ’ sp.* — 34·3 (3) — 61·1 (1) — 33·3 (1) — 67·2 (3) —
‘ Puerto Narino ’ sp.* — 26·6 (3) — — — — — 79·6 (3) —
‘ Rio Branco ’ sp.* — 37·8 (3) — — — — — 67·4 (3) —
‘ Rotpunkt ’ sp.* 38·4 (2) 25·9 (3) — — 50·9 (3) 36·0 (2) 84·9 (2) 77·3 (1) 62·2 (2)
‘ Smaragd ’ sp.* 66·7 (2) — 62·6 (2) 76·2 (2) 75·9 (1) 61·5 (1) 75·6 (2) — 84·9 (2)
Poecilia (Limia)
melanogaster Seegers 46 (15) 41 (15) 29 (10) 59 (12) 49 (14) 38 (25) 71 (10) 67 (18) 58 (17)
Pseudocrenilabrus
m. victoriae (Günther) 48·3 (29) 50·7 (35) 50·3 (29) 47·7 (34) 53·9 (43) 49·1 (55) 51·6 (19) 46·4 (12) 52·5 (17)

Figures in parentheses indicate number of broods.

*Undescribed species. Names used in German aquarium literature.
    717

T II. Statistical significance of correlation from multivariate

correlation analyses (two-way ANOVA) of sex ratio and temper-
ature (T ), pH, and temperature#pH in Apistogramma-species

Species P (T ) P (pH) P (T#pH) ÷2 d.f.

A. borellii 0·000924 0·055775 0·791684 2·690 3

A. caetei 0·496003 0·000000 0·303825 15·282 6
A. eunotus 0·000001 0·000796 0·168134 2·566 2
A. gephyra 0·000000 0·000001 0·036653 9·665 4
A. hongsloi 0·000000 0·000000 0·000002 15·103 4
A. meinkeni 0·004540 0·061353 0·361128 2·884 1
A. nijsseni 0·000000 0·000000 0·003309 75·714 8

caught in the wild by the subspecies-describer. Poecilia melanogaster used in our

investigations were descended from a laboratory-stock which was bred for about 25
generations in captivity.
All fish were bred in the laboratory at constant water conditions. Temperature, pH,
and water hardness were measured electronically. Deviation in temperature was less than
&0·2) C, deviation of pH less than &0·5. Water was purified by filtration and by weekly
changes of 30% of water. If water was changed it was brought to the experimental
conditions in advance. Manipulation of pH was established by humic acid, sodium
biphosphate and sodium bicarbonate.
For the experiments, pairs as well as groups of five males and 25 females were bred in
150#50#40 cm tanks. No statistical differences of sex ratio were found between single
pairs and groups, for instance neither between 15 different pairs of Apistogramma nijsseni
Kullander nor between four different groups of A. linkei Koslowski. Spawning in
artificial caves (46#30 mm, transparent boxes) was observed continually during the
spawning period of about 48 h in temperature reversal experiments or at least daily in
standard experiments by one of the authors (UR). The spawning period was regulated by
feeding the animals ad libitum with nauplii of brine shrimps (Artemia sp.). The numbers
of eggs, larvae or fry were counted regularly. Females and clutches were transferred from
the spawning tank to rearing tanks after fertilization. Females were removed from their
brood after the offspring became free swimming, which was usually after 7 to 10 days
from hatching.
Fish were bred and raised at 23, 26 and 29) C, and pH 4·5, 5·5 and 6·5, respectively.
Though different pH-schemes could not be investigated in all cases, we present all our
data of environmental sex determination (ESD) by pH, as there is strong evidence that
the sex of a variety of Apistogramma-species is influenced both by temperature and pH
(Table I). In general, clutches were bred at pH 5·5 as losses of eggs, larvae or fry were
lowest at this pH. Our investigations of the influence of temperature were focused mainly
on A. borellii (Regan), A. nijsseni, A. trifasciata (Eigenmann & Kennedy), and A. caetei
Kullander. To determine at what stage of development ESD took place, eggs, larvae, and
fry were transferred at different times from 23 to 29) C, and the reverse. The moment the
last egg of a clutch was fertilized we chose as time zero. The maximum duration of a
spawning we observed was 1 h, therefore differences in age of eggs within a clutch (error)
do not exceed this time in our experiments. Sex was determined according to sexually
dimorphic characteristics which were described by Kullander (1980, 1986).
For statistical purposes, experiments were used only if the total loss of eggs and
offspring within a brood was less than 10%. Therefore, differential mortality did not
significantly affect the results. The CSS-Statistica program was used for statistical
calculations. Data for all temperature/pH-combinations are not yet available for all
species, and only the data of seven Apistogramma-species were examined by multivariate
correlation analysis (two-way ANOVA) which can handle unbalanced data (Table II). In
addition, the correlation between temperature, pH, and sex ratio for all species examined
718 . ̈  . 

T III. Spearman rank correlation for temperature and pH

Offspring/brood
Species R (T ) P (T ) R (pH) P (pH)
mean min max n

Apistogramma
agassizii 85·6 51 133 16 0·89 <0·001 0·00 1·0
borellii 72·2 25 122 17 0·79 <0·001 "0·52 0·14
cacatuoides 68·5 34 97 25 0·60 <0·001 "0·72 <0·001
caetei 77·8 19 174 65 "0·35 <0·004 "0·11 <0·37
diplotaenia 59·8 31 81 11 0·89 <0·001 "0·29 0·37
eunotus 113·4 56 155 18 0·87 <0·001 "0·37 0·12
geisleri 62·0 33 87 7 0·94 0·001 — —
gephyra 88·6 43 143 27 0·76 <0·001 "0·50 <0·008
gibbiceps 71·3 42 109 15 0·93 <0·001 "0·33 <0·22
gossei 62·3 49 73 6 0·95 <0·001 — —
hippolytae 82·7 62 114 7 0·84 <0·016 "0·15 0·73
hoignei 55·9 42 69 7 0·86 <0·012 — —
hongsloi 52·2 15 90 28 0·70 <0·001 "0·41 0·028
inconspicua 40·5 24 58 8 0·86 <0·030 — —
iniridae 77·2 70 83 5 0·86 <0·058 — —
linkei 41·8 19 68 50 0·93 <0·001 "0·24 0·087
luelingi 76·5 54 93 4 0·89 <0·106 — —
macmasteri 90·2 73 117 6 0·95 <0·003 — —
meinkeni 65·0 47 83 12 0·91 <0·001 "0·30 0·33
mendezi 75·8 54 97 4 0·89 <0·106 — —
nijsseni 58·5 24 121 150 0·92 <0·001 "0·19 0·015
norberti 95·9 26 215 15 0·83 <0·001 "0·33 0·22
ortmanni 48·4 41 57 5 0·94 <0·014 — —
paucisquamis 74·6 59 116 11 0·87 <0·001 "0·36 0·27
pertensis 37·4 19 59 12 0·86 <0·001 — —
resticulosa 63·2 37 88 10 0·81 <0·004 "0·49 0·14
staecki 51·8 31 67 6 0·95 <0·001 — —
steindachneri 89·5 51 141 12 0·86 <0·001 "0·56 <0·057
trifasciata 46·2 22 80 45 0·94 <0·001 — —
uaupesi 48·3 29 80 16 0·89 <0·001 "0·42 0·10
‘ Breitbinden ’ sp.* 62·2 29 100 15 0·86 <0·001 "0·17 0·53
‘ Gelbwangen ’ sp.* 59·4 35 78 5 0·86 <0·058 — —
‘ Orangeschwanz ’ sp.* 153·9 111 191 8 0·88 <0·004 "0·21 0·60
‘ Puerto Narino ’ sp.* 86·8 67 122 6 0·87 0·021 — —
‘ Rio Branco ’ sp.* 68·7 55 88 6 0·87 0·021 — —
‘ Rotpunkt ’ sp.* 79·9 36 151 15 0·88 <0·001 "0·14 0·60
‘ Smaragd ’ sp.* 78·8 23 181 12 0·73 <0·001 "0·02 0·93
Poecilia (Limia)
melanogaster 28·4 12 45 136 0·79 0·011 "0·57 0·10
Pseudocrenilabrus
m. victoriae 25·2 6 41 273 0·15 0·68 0·10 0·78

Offspring/brood: mean: mean of all broods of a species; min and max: minimal and maximal absolute
number of offspring within a single brood; R (T ): Spearman rank correlation coefficient for temperature;
R (pH): Spearman rank correlation coefficient for acidity; P (T ) and P (pH): level of significance.
*Undescribed species. Names used in German aquarium literature.
    719

was tested using the Spearman rank correlation coefficient, especially designed for smaller
samples of non-parametric data (Sachs, 1984).
In most temperature schemes the influence of pH on the sex ratio of the offspring was
tested. For estimation of the temperature effect on sex ratio all data from different
pH-values within each temperature scheme were pooled before being tested using
Spearman’s method (Table III).
All experiments have been carried out in accordance with the national law. Voucher
specimens are deposited at the Zoologisches Forschungsinstitut and Museum Alexander
Koenig, Bonn, Germany.

RESULTS
In Apistogramma borellii, A. eunotus Kullander, A. gephyra Kullander, A.
hongsloi Kullander, A. meinkeni Kullander and A. nijsseni temperature was the
significant dominating factor affecting sex ratio. Whereas in A. caetei there was
no statistically significant impact of temperature on the sex ratio which was,
however, strongly influenced by pH (Table II; Fig. 1). Though not statistically
significant, there seemed to be a tendency in A. caetei to produce relatively higher
rates of males at 23 than at 29) C, which would be the reverse of observations in
other species.
In the Apistogramma-species, for which data have been analysed by multi-
variate correlation analysis, some influence of pH on the sex ratio could be seen
(Table II; Fig. 1). The lower levels of significance in A. borellii, A. eunotus and
A. meinkeni may be a result of the relatively small data sets in these species.
In total, the correlation between temperature and sex ratio of offspring of 33
species of Apistogramma was significant (Table III). For example, the sex ratio
of offspring in A. trifasciata at 23) C was skewed towards females, at 29) C
towards males, and at 26) C the number of males and females was approximately
balanced (Fig. 2). Four other species of Apistogramma seemed to show the same
response to temperature, but significance could not be shown since only four or
five broods could be raised for this investigation. Poecilia melanogaster showed
the same response to temperature as most species of Apistogramma, at 23) C the
sex ratio of offspring was skewed towards females, at 29) C towards males, and
at 26) C the sex ratio was again approximately balanced, whereas neither
temperature nor pH influenced the sex ratio of offspring in Pseudocrenilabrus m.
victoriae (Table III).
To determine the stage of development at which temperature influenced the
sex ratio in Apistogramma, eggs, larvae or fry of A. trifasciata were transferred at
different developmental stages from a male-determining temperature (23) C) to a
female-determining temperature (29) C) [Fig. 3(a)], and the reverse [Fig. 3(b)].
Both experiments revealed that the influence of temperature depends on the time
after which the brood is transferred. Broods transferred 800 h or later from the
end of spawning (time zero) had sex ratios characteristic of the initial tempera-
ture. Those transferred within 0 to 72 h had sex ratios characteristic of the final
temperature. Change in sex ratio was linear for broods transferred at later
intervals up to 800 h, indicating that temperature effects are cumulative during
the first 800 h of development.
According to Rubin (1985) pH affects the sex ratio in Apistogramma borellii
and A. caucatoides Hoedeman (which is probably a spelling mistake for
100 100 100 100 100 100 100
(a) (b) (c) (d) (e) (f) (g)

50 50 50 50 50 50 50

T
0 0 0 0 0 0 0
23 26 29 23 26 29 23 26 29 23 26 29 23 26 29 23 26 29 23 26 29
100 100 100 100 100 100 100

50 50 50 50 50 50 50

pH
0 0 0 0 0 0 0
4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5
100 100 100 100 100 100 100

50 50 50 50 50 50 50

T × pH
0 0 0 0 0 0 0
4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5 4.5 5.5 6.5
F. 1. Percentage of males dependent on temperature (T), top; pH, centre and temperature and pH (T#pH) lower in Apistogramma-species. (a) A. borellii;
(b) A. caetei; (c) A. eunotus; (d) A. gephyra; (e) A. hongsloi; (f) A. meinkeni; (g) A. nijsseni. T#pH: 1=29, ,=26 and #=23) C.
    721

100

80

60
Male (%)

40

20

0
23 26 29
Temperature (°C)
F. 2. Influence of temperature on sex ratio in Apistogramma trifasciata. n=15 at each temperature.
Mean; standard error ([ ]); standard deviation (T).

A. cacatuoides Hoedemann in Rubin’s article). Therefore we investigated also

the influence of pH on a variety of Apistogramma-species. In some but not all
Apistogramma-species the sex ratio of offspring was biased significantly by pH
(Tables I, III). The influence of pH is quite different from that of temperature.
In A. nijsseni for instance the influence of temperature seemed to be the
dominant factor [Fig. 4(a)], whereas in A. borellii both, high temperature and low
pH level, revealed a high percentage of males [Fig. 4(b)].

DISCUSSION
Whereas genotypic sex determination has been shown in many fish species
(Price, 1984), ESD in fish has been shown so far in only a few (Conover &
Kynard, 1981; Rubin, 1985; Sullivan & Schultz, 1986; Middaugh & Hemmer,
1987; Schultz, 1993). Our experiments prove that sex is determined in most, if
not all, Apistogramma-species and in Poecilia melanogaster by environmental
factors such as temperature and pH, and ESD in fish may be more common than
currently appreciated.
In contrast to temperature-dependent ESD in reptiles which typically exhibit
predominately all male or all female sex ratios over a wide range of incubation
temperatures (e.g. Ferguson & Joanen, 1982; Standora & Spotila, 1985; Bull
et al., 1990; Wibbels et al., 1991; Viets et al., 1993), in Apistogramma-species, as
in Menidia menidia (Conover & Heins, 1987a, b), ESD by temperature is not an
‘ all-or-none ’ effect. If incubation temperature during the first days after
fertilization is changed from 26) C, a temperature which yields a balanced sex
ratio, to a warmer (29) C) or cooler (23) C) temperature, respectively, sex ratios
in offspring of Apistogramma shift in both cases about the same amount, but in
opposite directions. Thus, we cannot find differences in sex determining poten-
cies with regard to the different temperatures we tested. Otherwise unbalanced
changes of sex ratios should be expected under these conditions, as Bull et al.
(1990) found in turtles.
722 . ̈  . 

(a)
100

Sensitive
80 period

60
Male (%)

40

20

0
0 200 400 600 800 1000 1400
Time (h)

(b)
100

Sensitive
80 period

60
Male (%)

40

20

0
0 200 400 600 800 1000 1400
Time (h)
F. 3. (a) Percentage of males after transfer of eggs, larvae or fry of Apistogramma trifasciata at different
times from 23) C, at which temperature reveals a low percentage of males, to 29) C, at which
temperature reveals a high percentage of males. (b) Percentage of males after transfer of eggs,
larvae or fry of Apistogramma trifasciata at different times from 29) C, at which temperature
reveals a high percentage of males, to 23) C, at which temperature reveals a low percentage of
males. Mean; standard error ([ ]); standard deviation (T); arrow: expected percentage of males if
clutch is not transferred to lower/higher temperature.

The sex of progeny of Apistogramma-species is therefore not determined at

conception, but within a sensitive period extending from 0–35 days after
spawning. This agrees with findings that heteromorphic sex chromosomes are
absent in all neotropical cichlids (Kornfield, 1984).
Sex determination in the Atlantic silverside (M. menidia) is under control of
both genotype and temperature during a specific period of larval development
    723

100

80
Male (%)

60

40

20

6.5

5.5 29
pH
-le

26
(°C)
ve

ture
l

4.5 23 p e r a
Tem
(a)

100

80
Male (%)

60

40

20

6.5

29
pH

5.5
-le

26 (°C)
ve

ture
l

4.5 23 p e r a
Tem
(b)
F. 4. (a) Influence of temperature and pH-level on sex ratio in Apistogramma nijsseni. (b) Influence of
temperature and pH-level on sex ratio in Apistogramma borellii.
724 . ̈  . 

(Conover & Kynard, 1981; Conover, 1984). The hypothesis of ESD suggests
environmental determination of sex whenever the environment affects the
relative fitness of males and females differentially and when the environmental
conditions that offspring enter cannot be chosen (Charnov & Bull, 1977). In
addition, parental adjustment of sex ratio in offspring during the period of
care may affect the reproductive success of male and female offspring differ-
ently (Trivers & Willard, 1973). Adaptive variation in environmental and
genetic sex determination in M. menidia according to this hypothesis could be
demonstrated by Conover & Heins (1987a, b): populations at different latitudes
compensate for differences in thermal environments and seasonality by adjust-
ing the response of sex ratio to temperature, and by altering the level of
environmental control in contrast to genetic control. Large size, as the result
of a longer growing season, enhances the survival and fecundity of female
M. menidia, whereas males do not appear to be greatly affected by body size.
Thus, it is an advantage to populations in the south with long breeding
seasons to hatch females by ESD first, whereas populations in the north with
short breeding seasons do not.
Typical Apistogramma are small, shelter-breeding fish, living in the leaf litter of
small rivers and brooks. They are in most cases polygamous, but in rarer cases
monogamous. Females care for their brood for about 10–50 days. Until now,
there has been no demonstration of either any ecological significance of ESD or
any selective evolutionary advantage for the occurrence of ESD in
Apistogramma-species. But recent investigations in brooks of the Rio Negro
system (Brazil) by one of us (UR) revealed a temperature gradient with respect
to water depth, ranging from about 30) C or more near the surface to 23) C or
less at the bottom. No pH-gradient was found between different strata of the
water, but there was one at different times of the day. The temperature gradient
was correlated with different numbers of individuals and different percentages of
males (Römer, unpubl.), while no such effect was found in relation to the pH.
Thus, we presume that our findings of an ESD in the laboratory is of importance
in the wild, too. Preliminary field studies seemed to reveal a socially influenced
temperature scheme for ESD in Apistogramma, instead of a seasonally or
geographically influenced one, as found in Menidia, but further investigations are
necessary.
All investigated Apistogramma-species, except A. caetei, seemed to follow a
common pattern of control of the sex ratio, especially in regard to temperature
(Fig. 1), whereas A. caetei seemed to show an inverse response of sex ratio to
temperature and additionally, a high response to the pH-level. No reason for
this can be found in experimental design nor in available ecological data. At
present we can only speculate that differences in habitat distribution, com-
pared to other Apistogramma-species investigated by us, may cause the
effect. A. caetei is found in streams and brooks in the vicinity of Belém while
all the other species originated from the Amazon, Orinoco or Paraguay Basins.
A. caetei, as well as two additional Apistogramma-species which were not
available until now, represent the most eastern forms of the genus from the
Brazilian Shield. This may result in different ecological conditions and
differing temperature and pH regimes, leading to an altered ESD system in
A. caetei.
    725

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