〔水SUISANZOSHOKU産 増 殖 44巻2号 (1996-H8)〕125-131

Food Consumption and Growth of dactylomela Rang ( : )

Stefanos PALATZIDIS1), Shigehisa YAMASAKI1), Hachiro HIRATA2), and Takehiko IMAI1)

1) Faculty of Fisheries , Kagoshima University, Shimoarata 4-50-20, Kagoshima 890, Japan 2) Faculty of Agriculture , Kinki University, 3327-204 Nakamachi, Nara 631, Japan

Abstract The consumption of two macrophytic algae, namely Ulva pertusa and Enteromorphalinza, by the adult gastropoda Aplysia dactylomela was studied for a period of 36 days under laboratory conditions. were fed three times a day to satiation. Both algae were well consumed and promoted increase in the body weight, body length as well as the egg production of the gastropoda. The consumption of Ulva by Aplysia was 22.5 % of mean body weight eaten per day, and was independent of temperature, but Enteromorphaconsumption was 15.0 % and increased with increasing temperatures. Nevertheless, animals fed on Enteromorphahad a better growth rate and egg production than those fed on Ulva. The gross assimilation efficiency was highest on the algae giving the better growth (29 % for Enteromorphaand 14 % for Ulva). The study found that the detrimental thalli of Ulva pertusa and also that of Enteromorphamay eventually be controlled by the introduction of gastropods in fish or shrimp ponds.

A common phenomenon in Japanese sea water oxidation-reduction potential of the pond bottoms7). aquaculture, during late spring and early summer is Several authors have demonstrated that polyculture, the rapid growth of macrophyte algae, like Ulva and using herbivorous species to control algal blooms in Enteromorpha, when water quality parameters deterio- commercial fish ponds, is an effective way to improve rate. Initially the presence of the algae can improve the water quality8,9). Very few research have been con- quality of water through an uptake of the toxic ducted on the control of macrophytes in sea water metabolites. Many researchers have shown the remark- aquaculture. Some workers, however, have shown that able application of Ulva as a biofilter in fish and herbivorous gastropod abalone could be cultured using shrimp culture systems1-5). Nevertheless, uncontrolled Ulva lactuca as the main source of food, to convert algal growth, can lead to algal die-off, depleting D. 0. large quantities of rich algal biomass into a high value levels, and releasing large amounts of toxic metabolites marketable commodity10,11) in to the culture pond depressing fish growth and/or The opisthobranch mollusc Aplysia has been re- causing deaths). Shigueno observed that dead and ported as an efficient consumer of macrophytic algae, rotten Ulva sp. led to heavy pollution, reducing drama- and grows much more rapidly than most marine tically the growth of shrimp, due to a negative animals12). Although various reports have shown the

Received : March 22, 1995 Key words: Aplysia dactylomela, Macrophytic algae, Food consumption, Growth 126 S.Palatzidis et al. (1996) ability of Aplysia to consume Ulva and other mac- Stock of Ulva pertusa Kjellman was obtained from Hira- rophyte algae, few have studied the possibility of using kawa Beach, Kagoshima Prefecture. Collections were this in polycultural systems. Aplysia is a spe- made once a week, and the stock was renewed at the cies of potential economic value by itself, apart from same time. Algae were maintained outdoors, under the being utilized as a model preparation for neurophy- natural varying lighting and temperature conditions in siological and neurobehavioral studies, because of its four (4) 100-liter circular Plexiglas tanks, filled with well identified cells with known interconnections and natural seawater. The algae were grown unattached behavioral functions13,14) Furthermore, sea hares are and kept suspended in the water column by air diffus- of pharmacological interest and have attracted the ers situated at the bottom of each tank. attention of investigators working on chemical com- pounds of marine invertebrates. An interesting exam- Animal culture system ple is the characterization of the antibacterial and anti- Animals were acclimated for upto 1 week before neoplastic glycoproteins in Aplysia juliana, and A. being used for the experiment in two (2) 100 liter's kurodail5-16) Over and above this, in some countries outdoor Plexiglas tanks containing filtered and aerated eggs of sea hares are utilized as food. natural seawater, that was replaced daily. The thalli of The present work was conducted to study the poten- the green algae Ulva pertusa and Enteromorpha lima tial use of the Aplysia in polyculture systems with was offered as food and was available in great abund Ulva and Enteromorpha. Therefore, one of the dominant ance at all times. species of sea hares Aplysia dactylomela, found in the Three days prior to the experiment, the animals Kagoshima bay, Southern Japan, was cultured in were starved, for better determination and selection of laboratory conditions with the two macrophytae spe- the initial weights. Animals with average 211.1 ± 5.5 cies. Their ability to consume these two macrophytic g (range 199.7-219.8 g) live body weights, were di- algae, the improvement in body measurements, and egg vided into 6 groups of 3 individuals for each algae. production were observed over a period of 36 days. The approximately equal aggregate live weight groups of sea hares, were transferred into six (6) 45 l glass- Materials and Methods walled aquaria. Moreover they were divided into two Field collections dietary groups. The aquaria were housed in a room, Reproductively mature individuals of Aplysia dacty- and covered with opaque material to minimize the in- lomela Rang, body weights ranging between 170-180 g, fluence of light and assure favorable culture conditions were collected from the intertidal area close to the for the predominantly nocturnal sea hares. The sea Kamoike Marine Experimental Station of Kagoshima water in the aquaria was renewed every day, filtered University, Kagoshima Prefecture. Collections were through a 5 am mesh and stored for one day in 200 l made in mid-spring (March 1994) and represented the circular tanks before use. common body weights for Aplysia dactylomela in that Every 6 days the animals were weighed, after being season and area. placed on a paper towel for a few seconds to drain off The particular biotope, formed by concrete breakwa- excess water. Body length was measured at full exten- ter extends 1-2 km parallel to the shore, creating a sion every 12 days. For the measurements, the animals oblong, protected tide channel. Several conduits in the were allowed to crawl on a transparent plastic mate- wall of breakwater allow the water circulation with rial, forming a cylindrical tube forcing the animals to the sea. Depth of the water body along the channel move forwards. The tube diameter was adjusted to suit ranges up to 5 m. The animals were found in the habi- the size of each animal. This tube, with the animal in- tat covered by the green alga Enteromorpha linza. side was submerged in the water with one extreme Thalli of Enteromorpha linza J. Agardh, were col- leaning downwards. Measurements were taken with a lected from the same biotope habituated by Aplysia ruler inside the water, once the animal had extended dactylomela and that seemed to be the preferred food. the entire body length. Eggs were collected daily from Food consumption and growth of Aplysia 127

the holding aquaria, drained of excess water and weighed. A known amount of food was provided to satiation three times a day to each of the sea hares aquarium. The thalli of the algae given as food or un- eaten remnants each time was washed with tap water and weighed after centrifugation for 3 min in a con- verted washing machine to remove excess water. Dry weight equivalents of body weights, egg masses and seaweeds were obtained by correcting for moisture content. Moisture content was obtained by drying 18 or 30 separate samples of animals, egg masses and sea-

weeds at 110 °C and averaging the results. Fig. 1. Growth and egg production of Aplysia dacty- The water temperature during the 36 day period lomela fed on Enteromorpha linza and Ulva pertusa ranged from about 17 to 21 °C. This range of tempera- diets under laboratory conditions. Each point in the curve represents the mean •} S.D. of 9 anim- ture, depending on season, reflects the natural condi- als. Each column represents the average weight

tions for Aplysia dactylomela. Water temperature was of the egg masses deposited every day in each

monitored twice daily in each aquarium. Dissolved ox- aquarium. ygen (D.O.) was maintained close to air-saturation level with aeration. The salinity ranged from 36-37 Over the period of 36 days Ulva is consumed more ppt. The 36-day experimental period was long enough than Enteromorpha but overall growth on this alga is to double the average body weights in both treatments. only 73 % of that on Enteromorpha (Table 1). Feeding rates for each diet on a dry weight basis, were found Analysis of data to be 4.46 g/animal/day for Ulva, and 2.96 g/animal/ Treatments were compared using a one-way classi- day for Enteromorpha. For comparison, values including fication of ANOVA (F-test) and the Scheffe test was the dry weight of eggs produced as a measure of total used to detect the differences between the means. The growth, are presented in Table 1. The growth rate in- level of statistical significance was accepted as p < creased 0.27 g/animal/day, for the animals feeding on 0.05. Enteromorpha and 0.15 g/animal/day for those feeding on Ulva, based on eggs produced. The gross assimila- Results tion efficiency over the total study period, calculated Growth and food consumption on a dry weight basis, is given for A. dactylomela on The growth of Aplysia dactylomela, measured as dry each algal diet in Table 1. Enteromorpha appears to weight increase, achieved with the two species of sea- give the higher growth efficiency of 28.5%. However weeds is shown in Fig. 1. The growth data, including A. dactylomela feeding on Ulva appear to convert the dry weights of food consumed and eggs produced by food into body tissues at an efficiency of 13.7 %. On A. dactylomela fed on each alga, are summarized for the including the dry weight of eggs produced in the mea- entire study period in Table 1. Of the two seaweeds sure of total growth, the growth efficiency was ad- studied, significantly higher (p <0.01) rate of growth justed to 37.6 % for Aplysia eating Enteromorpha and was attained on Enteromorpha linza. The green alga 17.0 % for Aplysia eating Ulva (Table 1). Ulva pertusa, also supported comparatively good growth. Similarly, biometrics for the length, in Aplysia Egg production feeding on Enteromorpha was significantly higher (p < The amount of eggs produced by A. dactylomela feed- 0.05), than on a diet of Ulva. The values of length ing on each alga is given in Table 1. The dry weight of growth rate were found to be 0.25 cm when food was eggs produced per individual per day was 0.27 g for Enteromorpha, and 0.15 cm for Ulva. those eating Enteromorpha and 0.15 g for animals fed 128 S.Palatzidis et al. (1996)

Table 1. Growth characteristics, egg production and food consumption of Aplysia dactylomela fed on Enteromorpha linza and Ulva pertusa diets in laboratory conditions under a temperature regime 17-21 °C. Data expressed as mean ± SD

Differences were, *significant at p < 0.05; * *significant at p < 0.01. 1 Gross assimilation efficiency = growth (dry wt) X 100/ingested food (dry wt) . on Ulva (Table 1). The daily egg production of Aplysia the temperature and egg production was R = 0.768; p fed on Enteromorpha and Ulva is shown in Fig. 1. Some <0.01 for the animals eating Enteromorpha and R = periodicity that occurred in the egg laying behavior 0.565; p < 0.01 for those eating Ulva. during the experimental period, depending on exoge- Discussion nous or endogenous factors, is also presented in Fig. 1. The growth performance of the Aplysia in terms of Effect of temperature on consumption, and egg production body weight gain, length increase, egg production, and There was a strong positive correlation between the the consumption of the algae was significantly affected temperature and the consumption of Enteromorpha (R by the type of algae. While Enteromorpha linza prom- = 0.7; p <0.05); unlike in the case of Ulva. The trend oted a better biological performance of the gastropoda, was the same as noted previously in the values of cor- Ulva pertusa consumption (dry and wet weight basis) relation between food consumption and egg production; was significantly higher. Superior growth and egg pro- the highest correlation was for Aplysia eating Entero- duction among the animals fed on Enteromorpha may in- morpha (R = 0.742; p < 0.01). The correlation between dicate its higher nutritional valuel7), and explain the Food consumption and growth of Aplysia 129 preference for this alga observed in animals grazing consumption or egg laying in Aplysia species and also under natural conditions. Laboratory-grown A. dacty- in A. dactylomela, occur during the night29,31-33) Being lomela fed with Enteromorpha sp. had better growth a nocturnal species, Aplysia remains inactive during rate, egg production, and higher food consumption, the day light, and normally feeds only during hours of when compared to the Ulva fasciata17). darkness. In our experiment, the animals were kept Our results showed a similar trend except for the under dark conditions; the continuous regime enabled fact, that the consumption of Ulva pertusa was signifi- constant feeding and produced greater growth and egg cantly higher than that of the Enteromorpha linza. The production. This explains why we could obtain an in- lower consumption of Enteromorpha compared to Ulva, crease in body weight combined with egg production, can be attributed to a dramatic increase in egg produc- contrary to the earlier findings where a stagnation or tion, when the Aplysia was fed on this alga. While the loss of body weight was reported during the periods body weight gain of animals fed on Enteromorpha were when the adult animals were laying their eggsl2,34,35) 38 % higher than those fed on Ulva, egg production in- . The consumption of macrophytic algae by the gastro- creased by 84 %. Feed consumption is inhibited by the poda has been clearly demonstrated here and in other bag cell hormones that induce egg layingl8). Furth- works17,29) indicating that it could be utilized to con- ermore, the animals feeding on Enteromorpha, spends a trol the macrophytic swarm, especially that of the Ulva larger proportion of the time in laying and encapsulat- pertusa. Additional studies may consider employing ing the eggs and further packing the capsules into an other gastropoda species, and observe the improvement egg string, diminishing the time available for food of water quality parameters. These factors, combined consumption19). On the other hand, since Aplysia rapid- with the market for the animal either for neurophysiol- ly consumes the Ulva diet, digestion and absorption of ogy examinations or for the pharmacological industry, this alga may be lower20) . will determine the feasibility of its introduction in Consumption was positively correlated with temper- polyculture systems. ature when the food was Enteromorpha, but no correla- References tion was found between temperature and the consump- tion of Ulva. Egg production in both groups was 1) Vandermeulen, H. and H. Gordin (1990) : Ammo- affected by rise in temperature. This is in agreement nium uptake using Ulva (Chlorophyta) in inten- with the report of other authors who found that sive fishpond systems: mass culture and treatment temperature affected the oogenesis of some Aplysia spe- of effluent. J. Applied phycology,2, 363-374. cies due to increased activity of the bag cell which re- 2) Danakusumah, E. and H. 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ア メ フ ラ シ(AplysiadactylomelaRang)の 摂 餌 と 成 長

StefanosPalatzidisl)・ 山 崎 繁 久2)・ 平 田 八 郎3)・ 今 井 健 彦2)

(1)鹿児 島 大 学 大 学 院 連 合 農 学研 究 科,2)鹿 児 島 大学 水 産学 部,3)近 畿 大 学 農 学 部)

養 殖 漁 場等 の 栄 養 塩 類 の豊 富 な 場 所 で は,大 型 藻類 の ア ナ ア オサ(Ulvapeytusa)の 成 長 が 良 く,水 質 浄化 に大 き く貢 献 す る可 能 性 が 示 唆 され て い る。 しか し,現 在,ア ナ ア オサ の利 用 は その 検 討 が 始 ま っ た ば か りで 有 効 な利 用 法 の 開 発 が 待 た れ て い る。 そ こで,本 実験 で は,食 物 連 鎖 に基 づ い た ア ナ ア オ サ の利 用 に よ りア ナ ア オサ の 繁 殖 の 制 御 を行 う基礎 研 究 と して,大 型 藻類 食 性 の ア メ フ ラ シ の摂 餌 率 と 成 長 率 の 関係 を調 べ た。 なお,ア ナ アオ サ と比 較 す る た め に,ア ナ ア オ サ と同様 に魚 介 類 の 飼 育 槽 に繁 茂 しや す い ウ ス バ ア オ ノ リ(Enteromorpha lima)も 餌 と して用 い た 。 ア メ フ ラ シ に よ る ア ナ ア オサ の摂 餌 率 は,餌 お よ び動 物 体 の乾 燥 重 量 換 算 で22.5%/day,ウ ス バ ア オ ノ リで15.0%/dayと な り,ア ナ ア オ サ で特 に高 い値 を示 した。 そ の 摂 餌 率 は,ア ナ アオ サ の 場 合 は 温 度 に あ ま り左 右 さ れ なか っ たが,ウ スバ ア オ ノ リの場 合 は温 度 の 上 昇 に伴 って 増 加 した 。 また,飼 料 転 換 効 率 は,ア ナ ア オサ で は14%,ウ ス バ ア オ ノ リで は29%と な っ た。 これ らの 値 か ら明 らか な よ う にア メ フラ シ の成 長 が極 め て速 い こ と を 考 慮 す る と,ア ナ ア オサ に限 らず ウス バ ア オ ノ リ に関 して もア メ フ ラシ に よる そ れ らの藻 類 の繁 殖 の, 効 率 の 良 い制 御 が 期 待 され る。