HISTOLOGY GONAD MORPHOHROMATIALLY DEFINED OF …...histology gonad ased on. 5 morphohromatially-defined of gonadal 6 maturation stages aalone (haliotis squamata 7 reeve, 1846), in

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GSJ: Volume 7, Issue 7, July 2019, Online: ISSN 2320-9186 2 www.globalscientificjournal.com 3

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

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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

GSJ: Volume 7, Issue 7, July 2019 ISSN 2320-9186

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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

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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

Alejo-Plata, M. C., & Gómez-Márquez, J. L. 2015. Reproductive biology of 125

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Oaxaca, Mexico. American Malacological Bulletin, 33 (1): 1-12. 127

Awaji, M., & Hamano, K. 2004. Gonad formation, sex differentiation and gonad 128

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Hern´andez-Cruz, C. M. 2012. Effect of Macroalgae Protein Levels on 132

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Booth, A. J., & Weyl, O. L. F. 2000. Histological validation of gonadal 136

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African Zoology, 35 (2):223-231. 138

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reproductive biology of the Donkey’sear abalone, Haliotis asinina Linne. 140

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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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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

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Female Male


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Pre-proliferative Maturing Ripe Total spawning

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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

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Documents

HISTOLOGY GONAD MORPHOHROMATIALLY DEFINED OF …...histology gonad ased on. 5 morphohromatially-defined of gonadal 6 maturation stages aalone (haliotis squamata 7 reeve, 1846), in