Journal of Fish Biology (2001) 59, 11531163

doi:10.1006/jfbi.2001.1726, available online at http://www.idealibrary.com on

Comparison between micro- and macro-structure readings in

the age estimations of Cape hake
A. G*, N. R * F. D
*Centro de Estudios Avanzados de Blanes (CSIC), Cam de Sta. Barbara s/n, 17300
Blanes (Girona), Spain and Ministry of Fisheries and Marine Resources, National
Marine Information and Research Centre (NatMIRC), P.O. Box 912, Swakopmund,
Namibia
(Received 22 March 2001, Accepted 23 July 2001)
By counting otolith daily increments juvenile Cape hake Merluccius capensis were found to
grow faster than European hake Merluccius merluccius. The mean frequency of translucent
zones in juveniles 130240 mm LT were higher (32) than the annual frequency, although
their mean duration was shorter (1 month) than a season. Comparison between micro- and
macro-structure readings showed that the latter tended to overestimate the age of juvenile hake,
with 1 year old hake being overestimated by 18% in individuals of 160 and 210 mm LT.
 2001 The Fisheries Society of the British Isles

Key words: Cape hake; growth; otolith; first-year; Merluccius capensis.

INTRODUCTION
Fish otoliths commonly used for fish age estimation are a diary where
biological and environmental information can be recorded for each individual.
The daily increment technique was developed in the early 1970s (Pannella, 1971,
1974; Brothers et al., 1976), and since then the advancement of otolith daily
increment methodology is such that its use at present is common and essential to
understanding the early life history of fishes. In the last decade two major
collections of papers on otolith microstructure have been published (Stevenson &
Campana, 1992; Secor et al., 1995). According to Jones (1992), deposition of
daily increments appears to be a universal phenomenon under perhaps all but the
most severe conditions.
Analysis of otolith daily increments of larvae and juveniles provides direct
information on dierent aspects beyond an accurate determination of age, such
as duration of the larval period, time of hatching, metamorphosis and settlement,
growth and mortality. Therefore, indirectly it provides key information on the
dynamics of fish populations, such as the recruitment index (Rutherford et al.,
1997), or the eects of spawning date on survival and growth during the early
stages (Secor & Houde, 1995; Meekan & Fortier, 1996; Michaletz, 1997;
Narimatsu & Munhara, 1999). Moreover, it provides information on the early
juvenile school performance (Sakakura & Tsukamoto, 1997) and serves as an
instrument for larval drift and stock identification (Campana et al., 1989;
Thresher et al., 1989).
Author to whom correspondence should be addressed. Tel.: 34 972 336101; fax: 34 972 337806; email:
gordoa@ceab.csic.es
1153
00221112/01/111153+11 $35.00/0

 2001 The Fisheries Society of the British Isles

1154

. .

Recently, otolith microstructure has been used to determine whether the first
noticeable translucent zone in the macro-structure of otoliths can be considered
annual (Gordoa et al., 2000a). The reliability of the estimates of biological
parameters, critical for any study on the dynamics of fish populations, depends
on the precision of fish ageing (Eklund et al., 2000). It is particularly of
importance in fishes of commercial interest, in which stock assessment procedures are dependent on accurate age estimates.
Namibian hake stocks, jointly with those from South Africa, represent 33% of
the total world hake biomass (Pitcher & Alheit, 1995). This region, under the
influence of the Benguela upwelling, provides one of the worlds most important
fishing grounds. There are three species of hake in this region: Merluccius polli
Cadenat, which normally do not extend further south than Angolan waters and
are very scarce in Namibian commercial catches, the deep water hake Merluccius
paradoxus Franca and Cape hake Merluccius capensis Castelnau (Gordoa et al.,
1995). The latter is the dominant species in the commercial catch and is the
species selected for this study. A revision of age and growth of this species
(Lleonart & Morales, 1985) concluded that the large interannual variance of
growth parameters was caused by erroneous reading or basic faults in interpretation. One of the hake species with the greatest amount of literature on age and
growth is the European hake Merluccius merluccius (L.). Despite all these
studies, there is still no internationally agreed upon method for its ageing. The
diculty in interpreting European hake has been attributed to various factors.
Morales-Nin et al. (1998) concluded that European hake otoliths in the
Mediterranean did not lay down an interpretable ring pattern that would be
useful for age determination. These authors attributed the lack of interpretable
rings to the absence of a strong seasonal signal in the area inhabited by the fish.
In contrast, Namibian waters are characterized by a strong seasonal signal (Boyd
& Agenbag, 1985), which is reinforced because upwelling and solar heating
seasons virtually overlap (Gordoa et al., 2000b). Consequently, if environmental
seasonality is the major factor governing annual ring patterns in hake otoliths, it
should be expected to occur in Namibian hakes.
The age of juvenile M. capensis was investigated through counting daily
increments and measuring the frequency, date of formation and extension of the
major discontinuities in their otoliths (translucent zones, rings and nucleus).
Specific objectives of the research were to estimate the growth and the duration
of the first life stage of this species recorded in the nucleus, and to measure the
degree of concordance between macro and micro-structure readings for 1 year
hake.
MATERIALS AND METHODS
Most of the specimens used in this study were caught during the hake biomass survey
conducted, in most cases once a year, on board the RV Dr Fridtjof Nansen (October 1990
to February 1999). Some specimens were also collected from the commercial fishing
vessel Ribadeo. Sagittal otoliths were extracted, washed with tap water and stored dry in
paper envelopes labelled with biological information such as sex and total length (LT). A
total of 99 otoliths from fish ranging between 8 and 240 mm were examined in the present
study. Since the principal objective was the dating of the first annulus, most of the
samples corresponded to individuals >190 mm (Fig. 1), as from this size upwards the
presence of translucent zones was more frequent.

Number of otoliths

1155

14
13
12
11
10
9
8
7
6
5
4
3
2
1
0

60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250

LT (mm)
F. 1. Number of hake otoliths read for each length.

The macroscopic readings of the otoliths were performed on an image taken with a
binocular microscope by a digital camera connected to an image-analyser system. The
otoliths were mounted external side up on glass slides with Eukitt resin and viewed
against a dark background. Through macroscopic structure, translucent zones were
identified and characterized as dierent rings: annuli (seasonal growth variation in terms
of age), demersal (transition zone formed during the movement from pelagic to demersal
habitat), or false (translucent zones, which were apparently not associated with the two
factors mentioned above). All of the macroscopic features identified were marked over
the image so that they could be used as guidance for the subsequent process of
microscopic determination of daily growth increments, and for dating, to determine the
validity of the macroscopic readings as well as to determine the frequency, duration and
month of formation.
For microscopic examination, all of the selected otoliths were observed using a light
microscope connected to a digital camera and to an image-analyser system. The
transmitted light made the translucent zones appear bright. Otoliths were ground
following standard methods (Morales-Nin, 1992) along the frontal plane until the nuclear
area was exposed. This area extended from the first clearly visible increment bounding
the core to the last clearly visible increment within the growth discontinuity region and
was polygonal in shape (Fig. 2). After calculating the increment of this particular area
the polishing procedure continued and the next increments were counted along the
longest axis (nucleus-rostrum direction), because, being the thinnest part, it was the last
to be polished. When the increment were not clear, a prominent increment was followed
laterally to the closest clear increments sequence (Morales-Nin & Adelbert, 1997).
For locating the period of deposition and determining the duration of the translucent
zones, the month of hatching was calculated for each individual by subtracting the age in
months (determined by increment growth counts) from the month of capture. Then, by
adding the age at formation of translucent zones (determined also by means of increment
growth counts) to the month of hatching, the month at deposition could be obtained.
Finally, the degree of concordance between the macroscopic and microscopic techniques was estimated. The percentage of agreement between annuli determined by
macroscopic and microscopic techniques was examined. Each ring characterized as an
annulus was considered truthful if it started 15 days before or after 365 days. A margin
of error of 1 month was permitted, which is greater than that estimated from repeated
counts of the same otolith (c15 days). This error was measured over six otoliths for fish
of LT between 200 and 240 mm, on which more than one reading was taken, and the
dierences between them were c15 days.

1156

. .

F. 2. Silhouette of Cape hake otolith nuclear area: (a) first zone that corresponds to the pelagic period
and (b) second zone that corresponds to the transition period: from pelagic to demersal.

RESULTS AND DISCUSSION

The number of otoliths read successfully was 71, while 28 otoliths were lost
during the reading process. Most of the losses were due to overpolishing.
Otolith counts ranged from 104 to 708 increments in Cape hake of LT
80240 mm. As far as is known, this is the first study where such old individuals
(c. 2 years) have been determined by counting daily increments. There is no sign
of any loss of regularity in deposition of increments after the first year of life
(Morales-Nin, 1987). Nevertheless, there is the possibility of loss occurring at
older ages, since the extent of periodicity in increment deposition is traditionally
associated with physiological changes related to sexual maturity (Morales-Nin,
1987).
NUCLEUS

The nucleus of all otoliths analysed appeared as a polygonal structure similar

to that found in M. merluccius (Morales-Nin & Adelbert, 1997) and very similar
to that which can be observed in cod Gadus morhua L. (Lagarde`re et al., 1995)
and in bluefish Pomatomus saltrix (L.) (Hare & Cowen, 1997). This structure,
previously named as a primordium (Morales-Nin & Adelbert, 1997) is considered characteristic of gadids (Arneri & Morales-Nin, 2000). The primordium

1157

consists of two zones in Cape hake (Fig. 2): the first where the pattern of
increments is concentric and continues around a single first primordia, and the
second which starts with the formation of accessory growth centre and finishes
with the last growth increment originating from the first primordium. In this
study the mean duration of the two zones together was found to be 45 days, with
a coecient of variation of 155%. In M. merluccius of the Adriatic, the 70 days
duration of the two zones (Arneri & Morales-Nin, 2000) is appreciably greater
than that observed in this study. These authors consider that the first zone
corresponds to the pelagic phase (c. 40 days) and the second to the period of
transition to the demersal component (30 days). The dierences in the duration
of this period in the two species are substantial. These data suggest that the
growth of M. capensis during this period is faster than in M. merluccius. The
apparent rapid change to the demersal of this species could indicate that its
growth during this period is much faster than that of the European hake.
Significant monthly variations have not been observed in the duration of the
predemersal period of this species, despite the strong thermal seasonality of this
region. Water temperature is a fundamental controlling factor of larval fish
growth (Campana & Hurley, 1989). If the finalization of the nucleus in the hake
coincides with completion of demersal settling (Arneri & Morales-Nin, 2000),
then the ring thus far accepted as the demersal ring in this species is invalidated.
In 28 of the 71 otoliths analysed, translucent zones characterized as demersal
rings were identified. The counting of increments revealed that these rings began
to form from a minimum of 134 up to a maximum of 373 days. Thus, it is
meaningless to identify these zones as demersal rings when, as mentioned above,
the demersal process ends at c. 70 days.
FORMATION OF TRANSLUCENT ZONES

The presence of translucent zones in the macrostructure began in individuals

of 130 mm. The frequency of translucent zones increased with LT, with a 90%
frequency in fish of 190 mm. In these larger specimens, the mean number of
macroscopically visible rings was 3.2 (..=13). This frequency is greater than
that expected by annual deposition and even greater than the biannual frequency
found in some tropical fishes (Yosef & Casselman, 1995).
The mean duration of these translucent zones varies from 3 to 113 days with
an average of 29 days, with a positively-biased distribution of frequencies (Fig.
3), in which a sharp reduction is observed in the frequency of the rings with
duration. This duration is much smaller than the 89 months considered the
general period for translucent zones in temperate latitudes (Beckman & Wilson,
1995).
The formation of the translucent rings may have originated from variations in
biotic or abiotic factors, thus their duration could be related to the month and
the age at which they were formed. However, there appears to be no relationship
between the duration of the rings and the age of the individual at the time of its
formation. Although the month of formation did not show any significant
pattern, this may have been due to the small sample size, since the mean monthly
duration of the translucent zones shows a potential seasonality (Fig. 4).
Maximum values occurred in June and July which correspond to the beginning
of winter in the southern hemisphere.

. .

1158
33
30
27
Frequency (n)

24
21
18
15
12
9
6
3
0

<=0
(10, 20)
(30, 40)
(50, 60)
(70, 80)
(90, 100) (110, 120)
(0, 10)
(20, 30)
(40, 50)
(60, 70)
(80, 90) (100, 110)
> 120

Duration of translucent rings (days)

F. 3. Frequency distribution of translucent zones by their duration.

80

Rings duration (days)

70
60
50
40
30
20
10
0
10

6
7
Month

10

11

12

F. 4. Duration of translucent zones depending on the month of the year when they were formed. ,
Mean; , 10 ..;
, 196 ..

GROWTH

The growth curve fitted to the juveniles of M. capensis explained 70% of the
variance [Fig. 5(a)]. A substantial amount of variability not explained by the
model could probably be attributed to the natural variability of the species itself
or to environmental factors. The variability of the species is high, since, for
example, the specimens analysed which were born in 1996 were mostly from the
same season (austral spring) and therefore grew under very similar environmental conditions. However, the variability observed in growth [Fig. 5(b)]
indicated a high intrinsic heterogeneity within the population.

1159

300
(a)
260
220
180
European hake

140
100
60

LT (mm)

20
20

European hake
100

200

300

400

500

600

700

800

250
240

(b) Dec

Nov

220

Sep

Nov

210

Oct
Nov

190

Aug

Oct

Nov

200

Oct

Nov

180
160
380

Aug

Sep

230

400

420

440

460
480
Age (days)

500

520

540

560

F. 5. Growth: (a) with all the otoliths read, otoliths without () and with () translucent zones; (b)
otoliths from individuals born in 1996; the labels show the estimated birth month. The line in (a)
was fitted by LT =15 Age0439.

Faster growth occurs in those otoliths which do not have translucent zones in
their macro-structure during the early stages of growth. From c. 320 days
onwards, all of the otoliths have translucent rings in their macro-structure and
the considerable variability in growth is reflected in their extension or duration
(Fig. 6). These results clearly confirm that the presence and duration of the rings
are associated with periods of slow growth (Morales-Nin, 1989). However, they
cannot be associated with a climatological season because the duration of these
rings is much shorter that a season and because their frequency is >1 year
1.
Despite the great variability observed in the growth of this species, the growth
appears to be greater than the estimated growth for Adriatic hake (Arneri &
Morales-Nin, 2000). At 1 year the Adriatic hake reach 160 mm LT, while in this
study Cape hake reach 200 mm. The dierences in growth may be due to the
dierences in the primary production of these regions, the typically oligotrophic
Mediterranean system and the highly productive waters of Namibia (Estrada &

. .

1160
750

Age (days)

650

550

450

350

250
20

20

60
100
140
180
Number of days within rings

220

260

F. 6. Variability in the age of Cape hake >300 days as a function of the number of days counted within
translucent zones. A regression line with 95% CL has been fitted (r=0672).

10.3% of underestimation
of the macro-structure
reading
33.3% of agreement
between readings

56.4% of
overestimation of the
macro-structure reading
F. 7. Proportion of agreement between macro-structure and micro-structure reading, and distribution
of over- and underestimation.

Marrase , 1987). The dierences observed in the growth of these species are
similar to the dierences found in the duration of the predemersal phase.
CONCORDANCE BETWEEN MACRO- AND MICRO-STRUCTURE
READINGS

Macro-structure readings considered 39 rings as annuli, but micro-structure

showed that only 33% of them were deposited at c. 1 year (Fig. 7). The errors in
the reading of the macro-structure present a clear bias towards overestimation,
because from the total disagreement (under- and overestimation) in 84% of the
cases the annuli were formed when the individuals were <1 year. The tendency

1161

to overestimate the age in these otoliths is fully justified by the frequency in the
formation of translucent zones, since this is >1 year
1.
Mean LT of the individuals of 1 year was 223 mm, with values ranging from
180 to 240 mm. This mean size is not very dierent from that obtained by
the macro-structure reading, which suggests that despite the discrepancy in the
readings the results ultimately do not dier substantially. However, 32% of the
specimens of 160210 cm were observed to have completed 1 year of growth
according to the counting of increments, while from macro-structure reading
50% had already completed 1 year of growth. This is explained by the extreme
dispersion observed along the age axis. This circumstance may falsify the age
structure of the populations because it is always estimated on the basis of the
length-age key, giving an overestimation of the real frequency of the age 1 year
class and an underestimation of the 0 year class by 18%.
The authors are grateful to P. Martin for his corrections to the English manuscript.
This work was sponsored by the Agencia Espan ola de Cooperacio n Internacional.

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