International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:13 No:01 31

Reproduction Aspects of Marine Worms Perinereis cultrifera (Grube, 1840) in Langgur Waters, Lesser Kei Islands, Southeast ,

Martha Rettob, Diana Arfiati, Luchman Hakim, Lawrence Lumingas

 Abstract— Marine worm class Polychaeta, Perinereis cultrifera I. INTRODUCTION (Grube, 1840), is used by many local residents in the Langgur SOUTHEAST Maluku regency geographically consists of two waters as food. But in the collection the fishermen capture them island groups namely the Kei and Aru islands. Ecologically, the before the spawn. The practices might decline in catches since Kei islands cluster was dominated by coral reefs of hard rocky the purpose of the worms to surface is to release gametes. substrate. The Kei islands consist of the Greater Kei Islands Therefore, the catching period needs to be regulated based on and the Lesser Kei Islands (Nuhuroa and Dullah Island). The bio-ecological aspects. The purpose of this study was to observe area of the Greater Kei Islands is 581 km² while area of the the dynamics of P. cultrifera worm catches, to analyze Lesser Kei Islands is 3,468 km² [1]. Langgur waters are located distribution of egg diameter, fecundity, and spawning peaks and in the Lesser Kei Islands with recede distance ± 50-300 m. to establish catching regulation. Sampling was carried out every One of biological aquatic resources in Langgur waters is month in June 2010 through May 2011 in the Langgur waters marine worms of Polychaeta class known by the locals as using a line transect sampling method along the shoreline. At "lawar" or in the Keian language is called "S'u". This marine any time of collection, samples were weighed to determine the worm (Perinereis cultrifera (Grube, 1840)) is in the Nereidae biomass, then 5% of the samples preserved in 50% alcohol solution. From the samples preserved were taken 10 and 40 family. The P. cultrifera worms can be found in clean and clear individuals respectively for egg diameter measurement and waters, algae covered rocky corals, hollow corals, and in fecundity determination. The highest biomass was obtained in undamaged reefs. September of 6346 g that indicated the spawning peak. The P. cultrifera worm is used by the Langgur community as one largest eggs were 110 μm and the highest fecundity of 27,000 of the popular traditional food for generations. Until now this eggs was obtained in October. The dynamics of the catch showed marine worm is used as the nutritious side dishes with a fairly that mature gametes can be found throughout the year. Juvenile complete composition of nutrients (Table I). individuals were available only from June to August. Thus, the The worms are also sold to supplement the family income correct closing period for catching the P. cultrifera worms would with an average price IDR 50,000/kg. With these advantages, P. be from June to October. cultrifera worm has a potency to be exploited and developed in order to meet the needs of human beings. For instance, it is Index Term— polychaeta, Perinereis cultrifera, bio-ecology, used as a raw material for food and pharmaceutical industries marine worm catching. such as supplement foods or fortified ingredients in food products [2]. The reproduction process of marine worms is very diverse and has almost every type of sexual and asexual reproductions. M. Rettob is with the State Fishery Polytechnics at , Jl. Jend. Asexual reproduction includes budding, parthenogenesis and Soedirman, Sathean 97611, South East Maluku, Indonesia (corresponding epitoky, supported by a soft body and a segmented body parts author, phone: +62-85331063938; e-mail: [email protected]). so easily separated. P. cultrifera marine worm also has sexual D. Arfiati is with the Faculty of Fishery and Marine Science, reproduction where gametes are produced by proliferation Brawijaya University, Jl. Veteran 2, Malang 65145, East Java, Indonesia. L. Hakim is with the Dept. of Biology, Faculty of Mathematics and (fertilization) of peritoneum cells. These cells are released into Natural Sciences, Brawijaya University, Jl. Veteran 2, Malang 65145, the coelomic cavity as primary gametocytes. Gamete formation East Java, Indonesia. can occur along the cavity of the body or only in certain areas L. Lumingas is with the Faculty of Fishery and Marine Science, Sam called reproductive segment. In Polychaeta the fertilization Ratulangi University, Jl. Kampus Kleak, Manado 95115, North Sulawesi, Indonesia. occurs outside the body, sperm and eggs are released into the

128805-1301-9494-IJBAS-IJENS @ February 2013 IJENS I J E N S International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:13 No:01 32 water simultaneously. Upon fertilization planktonic larvae will are iteroparous, sexual reproduction is often accompanied by be formed [3]. transitional epitokous forms. Epitoky consists of morphological T ABLE I changes in the Nereid, shape changes for swimming to the NUTRITIONAL CONTENTS OF P. CULTRIFERA WORM surface and conduct spawning [3]. Compositio Content Production of gametes during spawning requires a lot of n energy to gonad and body contractions since spawning may Protein 13.5% Fat 0.11% occur by way of the body wall cracking on the body segments Ash 2.40% that carry mature gametes. High energy also is needed, Water 76.71% especially in the epitokous species, because 75% of the total Ca 1814.45 energy is used for the production of gamete tissue and in P. ppm cultrifera it is 79% [4]. The life cycle of P. cultrifera species is Fe 21.65 ppm 3 years with 4 phases of egg development [5]. Essential Histidine 0.34% By the time the sexual cells are matured, P. cultrifera worms’ amino acids body is changing, such as enlarged eyes, the middle seta and Threonine 0.57% Tyrosine 0.34% parapodia are modified as swimming organs. In addition to Methionine 0.23% many living in the water, many worms swim to the surface at Valine 0.68% night to release eggs and sperm [6]. Phenylalanine 0.53% Catching the worms before spawning may result in the Isoleucine 0.62% decline in catches. Moreover, this worm has not been Leucine 0.81% cultivated due to lack of knowledge in reproductive aspect. Lysine 0.77% Therefore, it is necessary to study the biological aspects of Unsaturated palmitoleate acid (C16: 16 mg reproduction for reference in determining the period of catch fatty acids 1, n-7) oleic acid (C18: 1 n-9) 56 mg suspension and for providing data for P. cultrifera worm linoleic acid (C18: 2, n- farming. Thus, keeping the sustainability of the worm for 15 mg 6) human benefits. alpha-linolenic acid In this paper we discuss the dynamics of the P. cultrifera 14 mg (C18: 3, n-3) catches, the results of the analysis of the distribution of egg 11-eichosanoat acid 14 mg diameter, fecundity, and spawning peaks. The results of this (C20: 1, n-9) study are used to recommend worm fishing management for arachidonic acid (C20: 4, 164 mg their sustainability. n-6) EPA (C20:5, n-3) 58 mg DHA (C22:6, n-3) 21 mg II. RESEARCH METHOD AND MATTERS The sampling method employed was line transects, 700 m In general, sexual reproduction of marine worm is related to long, along the shoreline. There were two transects 10 m from age (months) and the tidal period. This is related to the release the shoreline and 20 m apart. 14 points in 100-m separations of epitoky (the segment that brings mature gametes) which were determined as sampling stations in each transect. only happens once a year. In this way Polychaeta can P. cultrifera worms were caught at night at 18.30 till 20.00 at reproduce in mass and abundance in the water. Epitoky the time of the new moon when the water begins to receding ± phenomenon of sexual reproduction involves the so-called 3-4 m from the shoreline and the water depth was ± 30-40 cm. "individual epitoke". In asexual reproduction it is called The catch was made once a month starting in June 2010 up to "atokous" through shape transformation of individual worms. May 2011 using a 2-L spherical fishing net. The catch was kept The newly formed individuals are photopositive and swim to in a container and weighed to determine the biomass. Then 5% the surface. The photonegative individuals are asexual and are of the samples were preserved in 50% alcohol solution. 10 in the bottom. There are two mechanisms of epitoky, epigamy worms were taken from the preserved samples for egg diameter and schizogamy. In epigamy all parts of the body turns into measurement and 40 worms were taken for fecundity epitoky, and in schizogamy only the posterior part of the calculation. sexually mature worms escape anteriorly. There are two main Egg diameter measurement began by opening the worm's reproductive systems in Polychaeta based on the spawning, body above the parapodia from the anterior to the posterior to the semelparous and iteroparous systems. In semelparous release their eggs. The eggs were put in a Petri dish containing species the spawning occurs once in lifetime, while in distilled water and then separated from the coelomic clot using iteroparous species the spawning can be observed several a pipette. Some 150 eggs were randomly taken each month. Egg times during their lifetime. Although many types of Polychaeta diameter was measured using an eyepiece micrometer on a

128805-1301-9494-IJBAS-IJENS @ February 2013 IJENS I J E N S International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:13 No:01 33 binocular microscope type XSZ-107BN No. 000807 at a 400× the synchronization between temperature, salinity, DO, fixed magnification. To test the difference between the average pH, the length of day, tides, and the moon period. This diameters of the eggs each month a statistical F-test analysis blooming showed that spawning peak occurs in September. (analysis of variance) was performed. The variance analysis is P. cultrifera worms moving into the surface in July to used to test the variations between groups and within groups October are adult worms with mature eggs [9]. This means that [7]. Should there be a difference further tests with LSD 95% the Polychaeta worms that left the holes in the low light were conducted to determine the pair average diameter of intensity are not only P. cultrifera worms with mature eggs, but different eggs. also juveniles of other marine worms that cannot be yet 40 worms from the preserved sample of each catch were identified. taken to calculate the number of eggs or. The samples were weighed and the body length was measured, after which the body of the worm was cut in similar procedures to the egg diameter measurement. The cleaned eggs were dried in ambient temperature for 30 minutes and then weighed to obtain the gonad weight. The eggs were randomly taken from the worms

to achieve a certain weight of the egg sample. The eggs were then placed into a container and added up to 100 mL of distilled Biomass (g) water and shaken, so that all eggs have the same opportunities to be taken without certain selection [8]. The eggs were observed under a binocular microscope type XSZ-107BN No. 000807 at a 40× magnification and counted with a hand tally Months of catching counter. Fecundity is calculated using a combination of Fig. 1. Dynamics of the biomass from the catches in Langgur waters. gravimetric method, volumetric, and direct calculation with the following formula:

GVX F  (1) Q with F = fecundity, G = gonad weight (g), V = volume of dilution (mL), X = the number of eggs per mL, and Q = weight of egg samples (g). Later, the suspension period of worm catching can be determined based on the model of the largest egg development, Fig. 2. The Perinereis cultrifera worms appearance on the surface of the highest fecundity and biomass as well as the highest peak Wearlilir waters in September. of spawning. .

III. RESULTS AND DISCUSSION B. P. cultrifera Egg Diameter in Langgur Waters A. Perinereis Cultrifera Worm Catch in Biomass During the catch period June 2010 – May 2011, the eggs Catching figures during the period of June 2010-May 2011 were dominated by small eggs (Fig. 3). The average diameter of (Fig. 1) show that the caught worms in January-April and the largest eggs (110 μm) was obtained in October. Distribution September-December consisted of P. cultrifera worm’s mature of the egg diameter ranged from 40-240 μm and the body length gametes. Catches in June-August consisted of juvenile and ranged from 19.04 to 38.72 mm. However, the statistical test unidentified worms and adult P. cultrifera with mature gametes. results did not show significance that there was no difference The average lowest (17 g) and the highest (6346 g) biomasses in the average diameter of the worm eggs at any time of the were obtained in March and September, respectively. catch. The low biomass occurs because catches were few and The average diameter of the largest eggs (110 μm) obtained macroalgae such as Padina, Ulva, Sargassum and Turbinaria in the Langgur waters was close to that (113 μm) obtained in were abundant in the bottom of the waters that inhibit the the Wearlilir waters [10]. In general, the P. cultrifera worms movement of worms to the surface. However, in September the catched in October were dominated by adult individuals with worms spread densely enough to cover the surface of the small body size and carry mature eggs that are not entirely water, thereby reducing the visibility to the bottom of the water homogeneous in size. This indicates that the eggs that were (Fig. 2). Blooming of P. cultrifera in the surface is an impact of morphologically measured originated from individuals that

128805-1301-9494-IJBAS-IJENS @ February 2013 IJENS I J E N S International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:13 No:01 34 have extracted their eggs. Therefore, large homogeneous eggs length, while 72% change in fecundity depended on other have been extracted leaving behind maturing egg cells [11]. The factors such as changes in the environment and food large eggs in October indicated supporting environmental availability [15]. The average fecundity of P. cultrifera factors that induced high reproductive activity such as rapid obtained was few compared with the results of previous yolk deposition during vitellogenesis, hydration, and the studies because of differences in body size. The body length of formation of oil grains. Thus, one can say that the variation of the P. cultrifera worm obtained in this study was smaller than egg diameter and its development depends on internal that that in the earlier study. The high or low fecundity of the conditions, such as nutrient reserves, and external conditions, organism depends on the condition of the waters in which the such as physico-chemistry factors of the environment and food organism lives [16]. Organisms living in less nutrient waters will [12, 13]. produce fewer eggs and otherwise those living in nutrient rich water will produce high fecundity. Individuals with high genetic diversity will have large component of fitness that includes growth, fecundity, viability, and resistance to environmental changes and stress [17].

umber of eggs

N

Month of catching

umber of eggs

N Fig. 3. Distribution of the diameter of the eggs in the Langgur waters during the capture period June 2010 - May 2011. The average diameter of mature eggs of P. cultrifera found in Algeria Beach was ± 250 μm [6] and 333 μm was the largest Month of catching diameter found in Wearlilir coastal waters [9]. The discrepancy in the egg diameter was assumed due to differences in the Fig. 4. The fecundity of P. cultrifera worms in Langgur waters. environmental conditions and food in the waters that have an impact on the formation and development of gametes. The D. Strategy on Catching Period Management for P. average egg diameter of this study was smaller compared to cultrifera those of Algerian coast and of the controlled container in the Based on the model of egg development and fecundity for laboratory. one year, the largest egg diameter of 110 μm and the highest In general, the egg will be homogeneous before spawning. fecundity of 27,000 eggs were acquired in October. Judging The females with maturing gonads, are gaining uniform size from the dynamics of the catch during the one year in which eggs. However, in young females, the egg size varies with small the highest catches in biomass in September, then September ones grow to large eggs at the end of their life cycle [11, 12, 14]. was defined as the spawning peak. The catch in June through Small eggs were produced by small individuals that are August made up of unidentified juvenile marine worm species regarded as the result of the formation of schizogamy epitoky. and adult P. cultrifera worms. Therefore, the recommended Therefore, if these small individuals move to the surface they suspension period for P. cultrifera worms catching is June to will bring small matured egg with them. October on the grounds that closure in June to August is to let the juvenile marine worms grow, in September is to allow the C. P. cultrifera Fecundity in Langgur Waters adult individuals to surface with high biomass to reproduce, Fecundity observations were made on 249 female worms and in October is to protect individuals carrying the eggs with taken from the 40 individuals from preserved samples. The the largest diameter and high fecundity. fecundity obtained ranged from 7785 to 27,003 eggs in the The purpose of setting the catching period is to preserve or average body length 26 mm (Fig. 4). conserve the P. cultrifera species for the next generation. Worm fecundity P. cultrifera 1,500-136,500 eggs and the Conservation concept in biology is an attempt to ensure the relationship between the fecundity with body length was not continuity of the existence of species, habitats and biological significant, 28% variation in fecundity was affected by the

128805-1301-9494-IJBAS-IJENS @ February 2013 IJENS I J E N S International Journal of Basic & Applied Sciences IJBAS-IJENS Vol:13 No:01 35 communities, and interactions between species, and the [13] J. S. Ryland and P. A. Tyler, Reproduction, Genetics and rd species and the ecosystem. Distributions of Marine Organisms: 23 European Marine Biology Symposium. Fredensborg, Denmark: Olsen & Olsen, 1989. IV. CONCLUSION [14] D. W. Golding, "Endocrine programmed development and Our study showed that the dynamics of the one-year catch reproduction in Nereis," Gen. Comp. Endocrinol., vol. 52, pp. consisted of unidentified juvenile marine worms and the adult 456-66, 1983. P. cultrifera worms carrying matured eggs with the highest [15] M. Rettob, "Hubungan Fekunditas dengan Panjang Standar biomass in September of 6346 g that is determined as spawning Cacing Laut Perinereis cultrifera (Grube, 1840) yang Tertangkap di Perairan Pantai Wearlilir Kepulauan Kei Kecil peak. P. cultrifera has the highest fecundity in October, with as Kabupaten Maluku Tenggara," J. Biotropikal Sains, vol. 6, many as 27,000 eggs and an average diameter of 110 μm. 2009 (in Indonesian). The closure of the catching location should be [16] T. B. Bagenal, Methods for Assessment of Fish Production in implemented from June to October. P. cultrifera worm farming Fresh Water. IPB Handbook Third Edition. London: Blackwell Scientific Publications, 1978. efforts needs to be done so as not to disrupt the continuity and [17] D. W. Ariani, Diversitas Genetik Cacing Lawar Perinereis preserving biodiversity in the Langgur waters. cultrifera (Grube, 1840) Di Kepulauan Kei Kecil Kabupaten Maluku Tenggara. Masters Thesis. Dept. of Biology, REFERENCES Brawijaya University. Malang. 2011 (in Indonesian). [1] E. A. Renjaan, "Penyediaan Data Dasar dan Perspektif Pengembangan Wilayah Pesisir, Laut, dan Pulau-Pulau Kecil di Kabupaten Maluku Tenggara," in Seminar Otonomi Pengelolaan Wilayah Pesisir, Laut, dan Pulau-Pulau Kecil Kabupaten Maluku Tenggara, 2002. [2] M. C. Latumahina, A. M. Tapotubun, and I. K. E. Savitri, "Studi Kandungan Nutrisi Laor," Pattimura University, Ambon, Fundamental Research Report. 2007 (in Indonesian). [3] R. S. K. Barnes, P. Calow, P. J. W. Olive, and D. W. Golding, The Invertebrates, A New Synthesis. London: Blackwell Scientific Publications, 1989. [4] C. Cassai and D. Prevedelli, "Reproductive effort, fecundity and energy allocation in two species of the genus Perinereis (Polychaeta: Nereididae)," Inver. Rep. Dev., vol. 34, pp. 125- 131, 1998. [5] J. C. Andries, "Endocrine and Environmental Control of Reproduction in Polychaeta," Can. J. Zool., vol. 79, pp. 254- 270, 2001. [6] A. Rouabah and P. Scaps, "Life Cycle and Population Dynamics of the Polychaete Perinereis cultrifera from the Algerian Mediterranean Coast," Mar. Ecol., vol. 24, pp. 85- 99, 2003. [7] B. Scherrer, Biostatisque. Boucherville, Canada: Gaetan Morin, 1984. [8] M. I. Effendie, Metoda Biologi Perikanan. Bogor: Yayasan Dewi Sri Cikuray, 1979 (in Indonesian). [9] M. Rettob, Studi Beberapa Aspek Reproduksi Cacing Laut Perinereis cultrifera (Grube, 1840) yang Tertangkap di Perairan Pantai Langgur Kepulauan Kei Kecil Kabupaten Maluku Tenggara. Masters Thesis. Graduate Program, Sam Ratulangi University. Manado. 2007 (in Indonesian). [10] M. Rettob, Aspek Bioekologis Cacing Lawar Perinereis cultrifera (Grube, 1840) dari Perairan Wearlilir dan Langgur Kepulauan Kei Kecil Kabupaten Maluku Tenggara. PhD Thesis. Graduate School of Fishery and Marine Science, Brawijaya University. Malang. 2012 (in Indonesian). [11] A. Fischer, "Control of oocyte differentiation in nereids (Annelida, Polychaeta): facts and ideas," Fortschr. Zool., vol. 29, pp. 227-245, 1984. [12] P. J. W. Olive and P. R. Garwood, "Gametogenic cycle and population structure of Nereis (Hedistc) diversicolor and Nereis (Nereis) pelagica from North East England," J. Mar. Biol. Ass. U.K., vol. 61, pp. 193-213, 1981.

128805-1301-9494-IJBAS-IJENS @ February 2013 IJENS I J E N S