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Sperm biology of artificially induced common carp, Cyprinus carpio (Linnaeus,

1758)

Article · January 2014

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 International Journal of Fisheries and Aquatic Studies 2014; 1(6): 27-31

ISSN: 2347-5129
IJFAS 2014; 1(6): 27-31
© 2013 IJFAS
www.fisheriesjournal.com
Sperm biology of artificially induced common carp,
Received: 06-05-2014 Cyprinus carpio (Linnaeus, 1758)
Accepted: 10-06-2014

M. Nahiduzzaman M. Nahiduzzaman, S. Akter, M. M. Hassan, A. K. M. Azad Shah, M. A. R.

(a)Department of Fisheries Biology
and Genetics, Bangladesh Agricultural Hossain
University, Mymensingh, Bangladesh
(b)WorldFish, CSISA-BD, ABSTRACT
Rangpur, Bangladesh Hatchery production of common carp seed has been practiced for several decades in Bangladesh but
information on sperm biology of induced broods is limited. The study aimed to determine the sperm biology
S. Akter artificially induced broods to improve the current hatchery management. Sperm volume, motility,
Department of Fisheries Biology and concentration and pH were 2.04±1.07 µl g-1 of fish, 93±3 %, 1.77±0.49×1010 cells ml-1 and 7.59±0.29,
Genetics, Bangladesh Agricultural respectively. There has been a substantial variation (p<0.05) in volume (µl g-1 of fish), concentration (×1010
University, Mymensingh, Bangladesh cells ml-1) and motility (%) among the fortnightly collected sperm samples. The motility (%) of the fresh
sperm was similar in all the activation media tested, however, sperm were motile for longer duration in 0.3%
M. M. Hassan
than in 0.2% NaCl, tap water and distilled water. Sperm biology of the induced broods would be useful in
(a) Department of Fisheries Biology
and Genetics, Hajee Mohammad
breeding programs advancing the development of aquaculture of the species.
Danesh Science and Technology Keywords: sperm biology, breeding, common carp, aquaculture.
University, Dinajpur, Bangladesh
(b) School of Biological Sciences,
Flinders University, Adelaide, 1. Introduction
Australia Common carp, Cyprinus carpio (Linnaeus, 1758) being one of the fastest growing fish species
contributing 10% of the global freshwater aquaculture production [1]. The species is
A. K. M. Azad Shah domesticated in lakes and ponds for aquaculture and the adaptability to a wider geographical
Department of Fisheries Technology,
location and temperature regime has contributed to the introduction of the species in many parts
Bangabandhu Sheikh Mujibur
Rahman Agricultural University, of Asia and Europe. Considering the potential of the common carp in small scale aquaculture in
Gazipur, Bangladesh homestead and eutrophic ponds the species was introduced in Bangladesh in 1960 from China
[2]
. The contribution of exotic carps is 28% of the total aquaculture production in Bangladesh in
M. A. R. Hossain which common carp is one of the leading species [3]. Although the species is considered invasive
Department of Fisheries Biology and
because of rubbing out pond dikes, however, the other attributes such as faster growth, disease
Genetics, Bangladesh Agricultural
University, Mymensingh, Bangladesh. resistance, adaptability to environmental fluctuations, short turnover time and suitability in
polyculture has made the species a profitable choice in extensive and semi-intensive
aquaculture.
In Bangladesh the seed production of common carp is entirely dependent induced gamete
collection by hormone administration in hatcheries [4]. Better understanding of sperm biology of
the species can improve the hatchery management efficiency of the species. In addition, sperm
biology is intrinsically crucial for short and long-term sperm storage, which could also improve
the current hatchery practice. Sperm biology provides a reasonable basis for developing a
strategy for maximizing the fertility of a fish and also the development of preservation protocols
[5]
.
Correspondence: The biophysical properties of sperm include its volume, concentration, pH and most importantly
Md. Nahiduzzaman percentages of motile sperm. These attributes are also important indicators of sperm quality and
Fisheries Scientist/Hub Manager artificial spawning. Usually sperm volume shows a seasonal variation in fish which is lowest at
WorldFish, Cereal Systems the onset that gradually reaches the highest during peak of spawning season [6]. Along with the
Initiative for South Asia (CSISA) in
seasonal variation, volume of sperm per unit fish body weight also varies with the age of
Bangladesh.
Rangpur Hub, House-46/2, Road-1, species. A single sperm is capable of fertilizing an egg, however, millions of sperm are
Parjaton purba Para Rangpur-5400, ejaculated by a male leading to sperm competition that ensure fertilization of eggs by the most
Bangladesh. competent sperm [7]. Therefore, concentration of sperm is crucial to maintaining sperm to egg
ratios during artificial spawning.

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International Journal of Fisheries and Aquatic Studies

Fish sperm are immotile inside the genital tract and after 14; Merck, Germany) immediately after collection.
collection an osmotic gradient of extracellular solvent induce Concentration was determined in triplicates using a
motility. Motility of sperm is activated by changing the haemocytometer and expressed as the number of sperm cells ×
external medium pH in many of the aquatic species [8], 1010 ml-1. Sperm were diluted 4,000-folds in distilled water,
although it is not as much crucial as osmolality in carps. pH of and a droplet of the diluted sperm was placed in a
the water which is added during artificial fertilization should haemacytometer (area of the smallest square = 1/400 mm2,
conform with the pH of seminal plasma. In freshwater fish, a depth 0.1 mm) for counting sperm. The number of
lower osmolality of extracellular medium (0-300 mOsmol l-1) spermatozoa in five large squares (area mm2) of the counting
compared to the osmolality of seminal plasma induce motility chamber was counted at 400 magnification.
whereas the opposite happens to marine species [9]. After
activation, motility of cyprinid sperm is actively motility from 2.3. Activation medium and sperm motility
few seconds to minutes in milli-Q water [10]. Therefore, sperm The motility of sperm samples was evaluated using a light
have to be in contact of the eggs soon after activation to ensure microscope (Novex K-range, Holland) at 400 magnifications.
reaching micropile of eggs. Although common carp is one the Sperm motility was estimated by adding 19 l of distilled
major contributor in aquaculture in many parts of the world, water (24 mOsmol kg-1) as activating medium to 1 l of fresh
including Bangladesh, information on sperm biology of sperm on a glass slide. The motility was observed within 3 to 4
artificially induced fish is limited. Therefore, the present study s after activation and was expressed as the percentage of
evaluated sperm biology of hormone administered common actively forward moving sperm out of the total. Sperm
carp to improve current hatchery management practice in spinning or vibrating in place were not considered to be
Bangladesh. motile. Four solutions (0.2% NaCl, 0.3% NaCl, distilled water
and tap water) with different osmolalities were tested for
2. Materials and Methods motility activation of the fresh sperm. Motility of sperm in
2.1. Rearing of broods each of the solutions was observed in triplicates to assess the
Broods were reared in brood-rearing earthen ponds (800 m2) motility (%), duration of motility, and to compare motility
of Fisheries Field Laboratory Complex, Bangladesh among solutions from a pool of five males.
Agricultural University (BAU) from July 2010-May 2011.
Water quality parameters including temperature (25.07±5.12 2.4. Statistical analysis
ºC), pH (7.69±0.32), dissolved oxygen (5.63±0.42 mg l -1) and Motility percentage was subjected to arcsine transformation
total alkalinity (86.34±21.57 mg l-1) of the ponds were prior to statistical analysis. One-way analysis of variance
recorded during the breeding season. The brood fish were fed (ANOVA) was carried out to determine the variation of sperm
with a commercial diet (35% protein; Paragon Feeds Limited, volume, motility percentage, concentration and pH. Pearson
Bangladesh) twice daily at 4–5% of their body weight in the correlation was used to relate sperm biology and body traits of
rearing period. the broods. The Duncan’s Multiple Range Test (DMRT) was
used to compare means at 0.05 significant levels. All values
2.2. Sperm collection and spermatological parameters were expressed as mean ± standard deviation (SD).
Ready to spawn broods were collected from the ponds and
conditioned for 12 hours in the hatchery tanks of the Faculty 3. Results
of Fisheries, BAU. The fish were injected with carp pituitary The weight of male (n = 43) ranged between 370 and 2000 g
supernatant at 2 mg kg-1 of body weight and released in the (mean±SD; 681.88±375.06 g) and total length ranged between
conditioning tank. After 6 h of hormone injection, the males 28 and 56 cm (35.43 ± 6.93 cm). Biology of common carp
were captured from the tank using a scoop net and were laid sperm was determined as sperm volume (µl g-1 of fish),
on foam to wipe the urogenital pore. Gentle pressure was motility (%), concentration (cell ml-1) and pH (Table 1).
applied through the abdomen to remove urine, water, gut Sperm volume was correlated with total length and weight of
exudates and mucus to avoid contamination. Sperm were the fish. Sperm volume was also positively correlated with
collected in glass vials by abdominal pressure and the vials sperm concentration and fresh sperm motility. Fish length and
were immediately placed at 4 °C. Ejaculated sperm volume weight were negatively correlated with sperm concentration
was determined by the measuring pipette and expressed as µl. (Table 2).
Sperm pH was determined with a pH indicator strips (pH: 0–

Table 1: A generalized summary of brood size and sperm biology of common carp.

Items Means ± SD Minimum Maximum

No. of fish 43 43 43
Total length (cm) 35.43 ± 6.93 28 56
Total weight (g) 681.88 ± 375.06 370 2000
Sperm volume (µl g-1) 2.04 ±1.07 0.68 4.67
Sperm pH 7.59 ± 0.29 7 8.4
Sperm concentration (×1010 ml-1) 1.77 ± 0.49 1 2.82
Sperm motility (%) 93 ± 3 85 95

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International Journal of Fisheries and Aquatic Studies

Table 2: Correlations between sperm biology with length and weight of broods.

Weight Sperm Sperm Sperm concentration Motility

(g) volume (µl g-1) pH (×1010 ml-1) (%)
Length (cm) 0.972** 0.634** -0.139 -0.396** -0.147
Weight (g) 0.594** -0.176 -0.359** -0.127
Sperm volume (µl g-1) -0.019 0.649** 0.504**
Sperm pH -0.17 -0.162
Sperm concentration 0.515**
(×1010 ml-1)
*p < 0.05, **p < 0.01

Spermatozoa showed a substantial variation in volume (µl g-1 the sampling fortnights (Table 3). Sperm pH did not show any
of fish) (F=2.520; p=0.033), concentration (×1010 ml-1) significant variation (F=0.345; p=0.927) among the sampling
(F=9.578; p=0.00) and motility (%) (F=7.209; p=0.00) among fortnights.

Table 3: Temporal variations in sperm volume (µl g-1), concentration (×1010 ml-1), pH and motility (%) in a spawning season of the
artificially induced common carp (Mean ± SD).

Fortnight* Sperm volume (µl g-1 ) Sperm concentration (×1010 ml-1) Sperm pH Sperm motility (%)
1 1.90 ± 0.09 bcd 1.44 ± 0.15b 7.60 ± 0.22 94 ± 2ab
2 1.86 ± 0.69bcde 2.28 ± 0.11 a 7.63 ± 0.25 95 ± 0a
3 2.92 ± 1.00ab 2.15 ± 0.32a 7.50 ± 0.00 94 ± 2a
4 3.64 ± 0.96a 2.02 ± 0.29a 7.73 ± 0.40 95 ± 0a
5 2.32 ± 1.01bc 1.99 ± 0.49 a 7.55 ± 0.15 95 ± 0a
6 1.70 ± 0.20 cde 1.42 ± 0.21b 7.67 ± 0.29 93 ± 3ab
7 0.96 ± 0.14de 1.42 ± 0.18b 7.70 ± 0.57 91 ± 4b
8 0.77 ± 0.09e 1.08 ± 0.11b 7.56 ± 0.48 88 ± 3c
*Fortnight 1-2: December; 3-4: January; 5-6: February; 7-8: March
Values with different letters within a column are significantly different (p < 0.05)

Percentage motility of the fresh sperm was almost same at the mOsmol kg-1) that retained motility for 118 s. Both tap water
beginning of activation in the four activating media with (31 mOsmol kg-1) and distilled water (24 mOsmol kg-1)
different osmolalities. However, 0.3% NaCl (96 mOsmol kg-1) produced poor motility duration (58 and 56 s) (Table 4).
yielded higher motility duration (141 s) than 0.2% NaCl (67

Table 4: Effect of activating media (0.3% NaCl, 0.2% NaCl, tap water and distilled water) at a range of osmolalities on sperm motility
(%) and motility duration of common carp.

Osmolality
Activation solution Motility (%) Motility duration (s)
(mOsmol kg-1)
0.3% NaCl 96 93 ± 3a 141 ± 5a
a
0.2% NaCl 67 93 ± 3 118 ± 6b
Tap water 31 92 ± 2a 58 ± 3c
Distilled water 24 93 ± 2a 56 ± 4c
Means with different letters within a column are significantly different (p < 0.05)

4. Discussion important parameter in hatchery management and it is highly

Several factors contribute to sperm biology, including age, variable depending on species, fish size and season [19]. Sperm
length and weight [11, 12], rearing conditions of brood [13] and volume and concentration of common carp were similar to
methods of spawning induction [14]. Sperm biology changes in other carp species including Gibelion catla, Labeo rohita,
a spawning season showing a gradual increase and decrease in Cirrhinus cirrhosus, L. calbasu and Hypophthalmichthys
motility percentage and concentrations at the beginning and molitrix [20, 21].
end of spawning [15, 16, 17, 18]. Sperm volume of common carp A change in pH of external medium is one of the sperm
was lower at the beginning of spawning season and it activating factors in aquatic species [22]. The pH of sperm was
gradually increased with the time indicating the temporal alkaline in common carp. There was no significant difference
variation of sperm volume in a spawning season. Sperm observed pH among different fortnight. The duration of sperm
concentration of common carp varied between the sampling motility in Petromyzon marinus decreased with an increase in
fortnights in which the highest concentration was found in the pH, but the percentage of motile cells did not change over the
middle of the spawning season. Sperm concentration is an pH range 6.0–9.0 [23]. Ingermann et al. [24] reported pH

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International Journal of Fisheries and Aquatic Studies

sensitivity of sperm in Acipenser transmontanus, and effects of dilution ratio ions and osmolality on sperm
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