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HISTOLOGY GONAD BASED ON 4
MORPHOCHROMATICALLY-DEFINED OF GONADAL 5
MATURATION STAGES ABALONE (HALIOTIS SQUAMATA 6
REEVE, 1846), IN BANTEN COASTAL, INDONESIA 7
SYAMSUL BACHRY1 , DEDY DURYADI SOLIHIN
1 RUDHY GUSTIANO
2, KADARWAN 8
SOEWARDI3, NURLISA A. BUTET
3 9
1Department of Biology, Faculty of Mathematics and Science, IPB 16680, Indonesia 10
2Institute for Freshwater Aquaculture and Fisheries Extension, 16151, Indonesia 11 3Department of Aquatic Resources Management, FPIK-IPB 16680, Indonesia 12
*E-mail: [email protected] 13 14
Abstract 15
The identification of the gonadal maturity stage in the specimen is very important 16
for determining the spawning season, this is because it can facilitate the process 17
of sustainable fisheries management such as abalone H. squamata. This study 18
aims to identify, histologically, gonads through morphochromatically-defined of 19
gonadal maturation. The results showed that four stages of gonadal development 20
in male and female abalone H. squamata were found where the four stages 21
included pre-proliferation, maturation, maturity, and consumption. Therefore 22
morphochromatic definitions can be used as a practical tool for identification of 23
gonad abalone H. squamata. 24
25 26
Introduction 27
Abalone is one type of marine mollusk that is of exotic and economical 28
value because the abalone shell is used for decoration and the meat is very 29
popular as one of the delicious and popular foods (Setyono, 2006). Abalone is 30
one of the commodities of marine products with demand in large quantities, 31
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especially Asian countries including Japan, China, Taiwan, Indonesia, and others 32
(Sales & Britz, 2001; Gordon & Cook, 2013). 33
Abalone is a gastropod that broadcast spawners (Takami & Kawamura, 34
2017). Abalone gonads can be observed through the left side of the shell by 35
lifting the legs and epipodium (Cox,1962; Setyono, 2006). The gonads lining the 36
digestive gland are called canonical appendage, which is known as 37
hepatogonadal complex (Velez-Arellano et al. 2015). Generally, females gonads 38
have a greenish color and males are beige in color (Sobhon et al. 1999; 39
Singhakaew et al. 2003; Hadijah et al., 2013; Roux et al. 2013). 40
Abalone size during gonadal maturation differs according to type, 41
geography, and water quality conditions where the abalone lives (Capinpin et al. 42
1998). Some sizes of abalone when first gonad mature include of H. midae from 43
the east coast of South Africa showing a shell size of 20-25 mm (Wood & Buxton, 44
1996), H. asinina from the waters of Tanakeke Island in South Sulawesi has a 45
shell length > 60 mm (Hadijah et al., 2013) other abalone species such as the 46
Haliotis type hannai disc that is kept in hatchery have a shell length of 30 mm 47
(Awaji & Hamano, 2004). Abalone in the subtropical region is different at the 48
beginning of gonadal maturity. The H. cracherodii species get a shell size of 140 49
mm (Del Proo, 1992). In addition, histological studies of gonadal development in 50
the adult phase have been carried out in various species (Hooker & Creese, 51
1995; Wood & Buxton, 1996; Capinpin et al. 1998; Setyono, 2004). However, 52
scientific information regarding the morphology and histology of gonads in 53
abalone H. squamata from Indonesia waters has not been widely reported. 54
Therefore, the purpose of this study was to identify the stages of gonadal 55
development through histological descriptions the morphochromatically-defined 56
of gonad abalone H. squamata on the coastal of Banten, Indonesia. 57
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Materials and Method 59
About 113 specimens were collected in February 2017. Specimens were 60
collected on the coastal of Banten, Indonesia (105º 53’630’ E, 06º 49, 87,4’ S). 61
Specimens are surgically removed from the hypogonadal complex. Each 62
hepatogonadal complex is photographed and examined to describe the color and 63
consistency of the gonads, and the size relative to the size of the 64
hepatopancreas. Male and female abalone gonads from several abalone 65
samples were fixed in 10% formalin solution and processed following standard 66
histological techniques: gonad specimens were inserted in paraffin, and 3-5 µm 67
thick sections were obtained and stained with hematoxylin-eosin (Humason, 68
1979). Criteria for determining the level of gonad maturity in H. squamata follow 69
(modification of Wood & Buxton 1996; Capinpin et al. 1998; Setyono, 2006, 70
Velez-Arellano et al. 2015). Sex determination of surviving abalone can be 71
observed visually by opening the side of the abalone shell. The color of the male 72
gonad is characterized by beige and female gonads characterized by green. 73
74
Result and Discussion 75
A total of 113 H. squamata specimens were examined, observations were 76
made based on gonad maturity in morphochromatic and histological features. 77
The gonadal characteristics found include four stages (Pre-proliferative, 78
development, maturation and total spawning) for the ovaries and testes (Table 1, 79
Figure 1 and Figure 2). Size of abalone H. squamata shell morphology found at 80
the time of observation was 4-7 cm. 81
Gonad maturation cycle in abalone occurs throughout the year. However, 82
abalone has its own image for the level of gonad maturity based on geography 83
and abalone living conditions. Thus the image of gonads can be used as a tool to 84
differentiate the stages of abalone gonad maturity based on the same level. 85
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Several studies that have explained the stage of gonadal maturity of the Haliotis 86
genus based on histology (Wood & Buxton, 1996; Capinpin et al. 1998; Bilbao et 87
al. 2012). 88
Based on the observation of macroscopic features, histological 89
characteristics generally showed an increase in the number of stable cells in the 90
process of gonadal maturation. Thus an increase in the size of the gonad is 91
easily observed visually. This is also seen in the size of the hepatopancreas 92
which has decreased, because of the presence of nutrients transported to the 93
gonad. The color of the gonad changes when the cells mature inside, this is 94
because it shows the development of various levels of gonadal maturity based on 95
macroscopic features. 96
Observations based on this description have been successfully used in fish 97
species such as Labeo cylindricus (Booth & Weyl, 2000), where macroscopic 98
patterns have also been used to see ovarian development in crustaceans such 99
as Callinectes danae (Zara et al. 2013), which has been explained in terms of the 100
main macroscopic relationship and histological structure of the oviductal gland 101
morphology that correlates well in the macroscopic evaluation of molluscs such 102
as Octopus hubbsorum (Alejo-Plata & Gómez-Márquez, 2015), where 103
macroscopic features have been described for various stages of maturation in 104
gastropods of Hexaplex trunculus (Elhasni et al. 2010). 105
Each species has a variety of different stages of gonadal maturity, this is in 106
accordance with environmental conditions. But results revealed in this study 107
indicate a practical and accurate way of identification in determining the stages of 108
gonadal maturation, without the need to do a long and time-consuming 109
histological analysis. 110
Conculsion 111
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Based on morphochromatic observations on gonad abalone H. squamata, four 112
stages of gonadal development were found including the pre-proliferation, 113
maturation, maturing and spent stages. In addition, the Criteria for TKG II (the 114
initial development stage) is a criterion that is mostly found in the observation of 115
male and female gonadal morphology in February 2017 in the waters of 116
Binuangeun, Banten Province. 117
Acknowledgment 118
This research was funded by Indonesian Government through Directorate 119
General of Higher Education (DIKTI), Ministry of Research, Technology and 120
Higher Education. We would like to thank for the Center for Research and 121
Development of Cultivation Sea from Gondol-Bali (CRDMAGB), fishermen and 122
collectors of abalon for their information. 123
References 124
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Booth, A. J., & Weyl, O. L. F. 2000. Histological validation of gonadal 136
macroscopic staging criteria for Labeo cylindricus (Pisces: Cyprinidae). 137
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Capinpin Jr, E. C., Encena II, V. C., & Bayona, N. C. 1998. Studies on the 139
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Cox, K. W. 1962. California Abalones, Family Haliotidae. The Resources Agency 142
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Wood, A. D. & Buxton, C. D. 1996. Aspects of the biology of the abalone Haliotis 186
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Table 1. Description stage of gonad maturity in H. squamata species 196
Pre-proliferative Description
Immature gonads are characterized by little or no germinal
epithelium between the outer epidermis and the digestive gland (Fig.
1A and 1B). Female gonad has a greenish color and male gonad by
beige (Fig. 2A and 2B).
Maturing Gonads enter the gametogenesis process. Histology analysis
revealed a number of oogonia began to associate with trabeculae
(Fig. 1C); in males, trabeculae begin to form with the attachment of
spermatogonia and spermatocytes; (Fig. 1D). Externally, this organ
is still soft and the base of the hepatopancreas has not been
surrounded by gonads. Gonad tissue covers the digestive gland 25%
to 50% of the cross-sectional area. Female gonad has a greenish
color while the male gonad by beige colored (Fig. 2C and 2D).
Ripe Gonad maturity fully contains gametes. Histology reveals female
gonads are almost full of mature oocytes (Gambar. 1E), while the
male gonad shows that the trabeculae have begun to be covered by
spermatozoa (Gambar. 1F). Female gonads show full ripe by
covering 90% of digestive organs, as well as male gonads showing
full development 85% envelop the digestive gland. While the gonadal
organ ends appear enlarged for each gonad. The ovary shows a
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bright green color, with the appearance of mature oocytes. Testicles
show a bright beige color, with a viscous fluid containing
spermatozoa (Fig. 2E and 2F).
Total spawning Gonads show evidence of gamete release (total spawning);
histological observations showed that ovaries containing mature
gametes were reduced and began to empty. Likewise with the
testicular segment visible around the tubules, where the
spermatozoa empty themselves, the lumen of the gonad is partially
destroyed and the trabecula folds (Fig. 1H and 1G). Morphologically,
the gonads look soft and inconsistent. Hepatogonad looks soft; the
color is opaque green in females, while males gonad display with
opaque beige color (Fig. 2H and 2G).
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200
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201 Figure 1. Histology images of morphochromatic differences in stage of 202
gonadal development in H. squamata: A) pre-proliferative female, B) pre-203
proliferative male, C) maturing female, D) maturing male, E) ripe female, 204
F) ripe male, G) total spawning female, H) total spawning male. 205
Hepatopancreas (Hp), Spermatozoa (Sz), Trabeculae (Tr), Oocytes (Oc), 206
Nucleus (N), Nucleolus (NL), female gonads filled with oocytes (Ro). 207
Hematoxylin-eosin technique, were photographed with an Olympus 208
microscope at a 10× magnification. 209
210
211
Female Male
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Pre-proliferative Maturing Ripe Total spawning
212
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Figure 2. Morphological features of various stages of maturation of the 214
gonads of Haliotis squamata: A) pre-proliferative female, B) pre-215
proliferative male, C) maturing female, D) maturing male, E) ripe female, 216
F) ripe male, G) total spawning female, H) total spawning male. 217
218
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