Effects of culture density and bottom area on growth and survival of the cuttlefish <Emphasis Type="Italic">Sepia</Emphasis><Emphasis Type="Italic">officinalis</Emphasis> (Linnaeus, 1758)

The effects of culture density and bottom areas on cuttlefish (Sepia officinalis) culture were studied. Cuttle fish were cultured under three experimental combinations of culture density and bottom area: (1) high density and small bottom surface area; (2) low density and large bottom area; (3) high density and large bottom area. Each experimental protocol was repeated in triplicate. Average weights at the end of the experiment were of 65.8 ± 5.8, 87.1 ± 5.6 and 78.7 ± 5.9 g for cuttlefish cultured under the conditions of protocol 1, 2, and 3, respectively; these differences were significant between each of the three groups. Total biomass increased faster (up to 7.5 kg per tank) under the high density/large bottom area conditions (protocol 3) due to the larger number of animals and low mortality in those tanks. Growth rates (%bw day⁻¹) were different between protocols, with growth rates of 2.1 ± 0.1, 3.0 ± 0.2, and 2.5 ± 0.1%bw day⁻¹ obtained for cuttlefish cultured according to protocol 1, 2, or 3, respectively. Average feeding rates were similar for the three groups—10.7 ± 0.8, 9.7 ± 2.0, and 9.6 ± 1.1%bw day⁻¹ for cuttlefish cultured according to protocols 1, 2, and 3, respectively, while food conversions (%) were different—21.5 ± 3.2, 32.4 ± 2.5, and 27.0 ± 1.1%bw day⁻¹, respectively. Total mortality was high in the high density/small bottom area tanks, 30%, while it was very low for the groups cultured under conditions of low and high density/large bottom area, 4%. Based on these results, we conclude that culture conditions that provide large bottom areas also provide good survival conditions and promote growth in comparison those with small bottom areas, even under conditions of lower culture densities.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Immunological effects of glucan and <Emphasis Type="Italic">Lactobacillus rhamnosus</Emphasis> GG,a probiotic bacterium,on Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> intestine with oral <Emphasis Type="Italic">Aeromonas</Emphasis> challenges

The present study histologically examined the effects of glucan-containing and Lactobacillus rhamnosus GG (LGG)-containing diets on intestinal damages inflicted on Nile tilapia by Aeromonas challenges. Tilapia were fed control, glucan, and LGG diets for 2 weeks and were subsequently challenged with Aeromonas. The intestines were then histologically examined at 1, 7, 14, and 21 days post-infection. Mortality following the challenge was lower for the fish fed the glucan and LGG diets. The intestines of these groups also showed increased inflammatory cell infiltration and reduced intestinal damage from Aeromonas. Moreover, inflammatory cell infiltration occurred more rapidly in the glucan-fed than in the LGG-fed fish following the challenge. Before the challenge, the dominant mucous cell was the acid type in all the tests. After the challenge, the main mucus cell type in the proximal intestine of the glucan-fed fish shifted to AB-PAS double-staining cells, while in the LGG-fed fish, it remained the acid type throughout the test period, and the number of double-staining cells was smaller than in the control fish after the challenge. Thus, the different mucous cell and inflammatory cell responses show that glucan and LGG might have different immunostimulative effects, although they both reduced the intestinal damage following Aeromonas challenges.     

The effect of stocking density on yield,growth, and survival of Asian river catfish (<Emphasis Type="Italic">Pangasius</Emphasis><Emphasis Type="Italic">bocourti</Emphasis> Sauvage, 1880) cultured in cages

Asian river catfish (Pangasius bocourti Sauvage, 1880) were cultured at five different stocking densities in cages (submerged volume 1 m³) suspended in a dugout pond from August to November 2009. Pangasius bocourti fingerlings (mean weight 27.09 ± 0.54 g) were stocked at densities of 12, 25, 50, 100, and 200 fish m⁻³. At the end of 3 months, the harvest weights (gross yields) were, respectively, 2.05 ± 0.30, 5.20 ± 0.31, 10.60 ± 0.42, 19.98 ± 0.78, and 42.37 ± 0.41 kg m⁻³. The mean fish weights among the stocking densities of 25, 50, 100, and 200 fish m⁻³ were not significantly different, but were significantly higher than that of the 12 fish m⁻³ density. The specific growth rates among high stocking densities of 50, 100, and 200 fish m⁻³ were not significantly different; however, they were significantly higher than those of the low stocking densities of 12 and 25 fish m⁻³. Asian river catfish performed poorly at the lowest density. The results indicate an initial lower stocking threshold for Asian river catfish of above 5.20 kg m⁻³. The Asian river catfish cultured in small cages placed in a pond reached the desirable market size (>200 g) within a 90-day grow-out period. The results show that the maximum yield for Asian river catfish during a 3-month production cycle was not reached.     

Demographic and competition studies on <Emphasis Type="Italic">Brachionus ibericus</Emphasis> and <Emphasis Type="Italic">Proales similis</Emphasis> in relation to salinity and algal (<Emphasis Type="Italic">Nannochloropsis oculata</Emphasis>) density

We isolated the rotifers Brachionus ibericus and Proales similis from the sediment of shrimp tanks and studied their individual demographic characters and competition between them at two food levels (0.25?×?10⁶, 1.00?×?10⁶ cells ml^?1 of Nannochloropsis oculata at 25 °C) and salinities ranging from 10 to 30‰. Our hypothesis was that growth rates would be higher with increasing food levels and salinities. Observations were taken twice a day for life table studies and daily once for population growth experiments. Using survivorship and fecundity data, we derived various life history variables. Although the average life span (7.6?±?0.4 days) and gross reproductive rate (33.8?±?2.9 neonate female^?1 day^?1) of B. ibericus were higher than those of P. similis (average life span 5.4?±?0.6 days and gross reproductive rate 13.0?±?0.6 neonate female^?1 day^?1), the population growth experiments showed that P. similis had higher r values (0.32?±?0.005 day^?1) than B. ibericus (0.23?±?0.002 day^?1) at 1.0?×?10⁶ cells ml^?1 of N. oculata. The rotifer P. similis was more adversely affected due to the presence of B. ibericus than vice versa. The data are important for developing techniques for a large-scale culture of these rotifers as food in aquaculture.     

Long-term effects of stocking density on survival,growth performance and marketable production of the sea urchin <Emphasis Type="Italic">Strongylocentrotus intermedius</Emphasis>

Sea urchins were stocked at a density of 15 (D15), 30 (D30), 45 (D45) and 60 (D60) urchins/cage (0.3 m long × 0.2 m wide × 0.4 m high) in a laboratory culture environment for 16 months. The wet body weight (BW) and test diameter growth were monitored at 2-month intervals during the experiment. At the conclusion of the experiment, the surviving sea urchins were counted and the gonad wet weight (GW) and gonad color were measured. Specific growth rate (SGR) of body weight, survival rate (SR), gonad index (GI), gonad color difference (ΔE ₀₀), coefficients of variation (CV) of BW, GW, GI and ΔE ₀₀, total biomass yield (TY) and total gonad yield (TGY) per cage were calculated. Two marketable yield variables, graded according to gonad index, i.e., marketable biomass yield (MY) and marketable gonad yield (MGY), were also calculated. Coefficient of variation of final body weight (FW) and final test diameter (FTD) of sea urchins increased as the stocking density increased, indicating the existence of adverse social interactions. These adverse social interactions detrimentally affected FW, FTD, SGR, GW and GI (P < 0.01). Although SR decreased with the increasing densities, no statistical significant difference was detected. Sea urchins at D15 had the lowest gonad color difference (ΔE ₀₀) (P < 0.05). However, statistically equal CV of ΔE ₀₀ indicates this density effect was not a result of adverse social interactions. TY and TGY increased with increased density and can be described by the following equations: TY = 84.18X ^0.64, R ² = 0.999 and TGY = 24.16X ^0.38, R ² = 0.979. However, the MY and MGY were not significantly different among stocking densities. The results of this study demonstrate that in intensive culture S. intermedius at low stocking density can achieve high growth rate, gonad index and desirable color without decreasing the marketable yield. Farmers should choose to culture S. intermedius at low stocking densities.     

Monitoring of <Emphasis Type="Italic">Francisella noatunensis</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis> in farmed Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) in Brazil

Francisella noatunensis orientalis is a bacterium that causes emerging bacteriosis in Nile tilapia (Oreochromis niloticus) in many parts of the world, including Brazil. It is a non-motile, Gram-negative, strictly aerobic, facultative intracellular coccobacillus. This species of bacteria is responsible for low to high mortality in fish farms, causing economic losses for fish farmers. This study aimed to detect the presence of F. noatunensis orientalis using qPCR (real-time polymerase chain reaction) and to describe lesions caused by the bacterium in O. niloticus in Brazilian aquaculture. For this purpose, 360 fish from six fish farms (30 per farm) were sampled at two time points (n = 180 per sampling). Necropsies and histopathology were performed for lesion observation, in addition to qPCR and sequencing for detection and identification of Francisella species. Environmental data were collected using a multiparameter sonde YSI EXO2. All measured limnological variables were within the optimum range for cultivation of Nile tilapia. The major lesions present were melanization of the skin, splenomegaly, granulomas, and inflammatory cell responses. The prevalence of francisellosis varied from 0 to 86.66% between time periods and fish farms analyzed, and an outbreak was observed during the second sampling period. This study describes the prevalence of francisellosis in O. niloticus and reports that the lesions found are not exclusively associated with this bacterial disease.     

Long-term effects of stocking density on somatic growth,gonad index and survival of the juvenile sea urchin, <Emphasis Type="Italic">Strongylocentrotus droebachiensis</Emphasis>

Juvenile Strongylocentrotus droebachiensis (average wet weight = 4.0 g) were held at three initial stocking densities (15, 30 and 60 animals per tray, which equates to 0.25, 0.5 and 1 kg sea urchins/m² surface area, respectively) under constant light and temperature conditions (LD 24:0 and 8.8°C) for 380 days. The somatic growth and survival of the urchins were monitored during this period, and the gonad index (GI) was measured at the conclusion of the experiment. The sea urchins in the two lowest stocking density groups had 100% survival after 380 days in contrast to the highest stocking density group, which only had 67% survival at the conclusion of the experiment. Somatic growth was significantly higher in the lowest density group than in the medium density group, and this in turn was significantly higher than in the high-density group. The injury occurrence was highest in the high-density group (20%), lower in the medium group (7%), and there were no injuries observed (0%) in the low density groups. There were no significant differences in GI between the groups at the end of the experiment. In conclusion, the present study showed that stocking density has significant effects on somatic growth and survival of juvenile green sea urchin, S. droebachiensis and should be taken into consideration when designing sea urchin holding systems.     

<Emphasis Type="Italic">Virgibacillus proomii</Emphasis> and <Emphasis Type="Italic">Bacillus mojavensis</Emphasis> as probiotics in sea bass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) larvae: effects on growth performance and digestive enzyme activities

This study examined the effects of two probiotics (Virgibacillus proomii and Bacillus mojavensis) on the digestive enzyme activity, survival and growth of Dicentrarchus labrax at various ontogenetic stages in three separate experiments. These probiotics were incorporated as single or mixed into fish feed for a period of 60 days. The growth parameters, proximate composition of whole body, digestive enzymes and gut microbiology were monitored at regular. The increments in length and weight and the survival were significantly higher (P < 0.05), and the values of food conversions were significantly lower (P < 0.05) in fishes fed the probiotic. The administration of V. proomii and B. mojavensis in diet resulted in an increase (P > 0.05) in body ash and protein content and in the specific activity of phosphatase alkaline and amylase in the digestive tract of all the fishes. V. proomii and B. mojavensis persisted in the fish intestine and in the feed in high numbers during the feeding period (group 1: 5.8 × 10⁴ CFU/ml, group 2: 9.6 × 10⁴ CFU/ml, and group 3: 9.8 × 10⁴ CFU/ml day 60). The two probiotics V. proomii and B. mojavensis were adequate for improved growth performance and survival and for healthy gut microenvironment of the host.     

Effects of salinity,stocking density,and algal density on growth and survival of Iwagaki oyster <Emphasis Type="Italic">Crassostrea nippona</Emphasis> larvae

To determine the optimal salinity, stocking density, and algal density for hatchery culture of the Iwagaki oyster Crassostrea nippona larvae, three experiments with salinities of 14, 18, 22, 26, 30, and 34 practical salinity unit (PSU); stocking densities of 0.5, 1, 2, 4, 8, and 12 larvae ml^?1; and algal densities of 10, 20, 40, and 100?×?10³ cells ml^?1 were designed, which included the developmental stages from newly hatched D-larvae to pediveligers. Results showed that larval growth of C. nippona was the fastest at a salinity of 26 PSU, and when salinity was adjusted to a level that was lower or higher than this salinity, survival and growth rate of larvae declined (P <?0.05), resulting both in a decreased mean shell length and a high mortality. Larval growth decreased significantly with increasing stocking density. Larvae reared at 4 larvae ml^?1 had the smallest shell length (198.9 μm) and lowest survival rate (7.9%), whereas larvae reared at 0.5 larvae ml^?1 had the largest shell length (245 μm) and highest survival rate (66.3%) on day 13. And the shell length of larvae reared at 0.5 and 1 larvae ml^?1 was significantly (P?<?0.05) larger than the values in other treatments, except those reared at 2 larvae ml^?1 (P?>?0.05). When feeding the single-algal diet of Isochrysis galbana (clone T-ISO), the shell length of larvae increased markedly as the algal density was increased. Larvae reared at the highest algal density (100?×?10³ cells ml^?1) had the largest mean shell length; however, under the conditions of our experiment, there was no significant difference (P?>?0.05) in growth and survival rates between the treatments at algal densities of 40?×?10³ and 100?×?10³ cells ml^?1. For a large-scale culture, based on the results of this study, a salinity of 26 PSU, stocking density of 0.5–1 larvae ml^?1, and algal density of 40?×?10³ cells ml^?1 are recommended for an early development of C. nippona.     

The use of alternative prey (crayfish, <Emphasis Type="Italic">Procambarus clarki</Emphasis>, and hake, <Emphasis Type="Italic">Merlucius gayi</Emphasis>) to culture <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier 1797)

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3.0 ± 0.7 kg) than those fed crayfish (2.4 ± 0.6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1.7 ± 0.3 and 1.2 ± 0.2 %BW day⁻¹, respectively). Average feeding rates for the experiment were not different, being 6.5 ± 0.9 and 7.5 ± 0.9 %BW day⁻¹, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42.4 ± 2.7%) for octopuses fed squid compared to the ones fed crayfish (23.9 ± 1.9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183.0 ± 242.7 g, 1,175.6 ± 240.1 g and 922.3 ± 160.1 g, respectively. Overall growth rates for the experiment were 1.9 ± 0.2 %BW day⁻¹, 1.9 ± 0.3 %BW day⁻¹ and 1.1 ± 0.3 g, respectively. Final weight and growth rates were never different (P > 0.05) between octopuses fed squid and hake, but were always higher (P < 0.05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4.6 ± 1.5, 4.2 ± 1.3 %BW day⁻¹ and 5.1 ± 0.9 %BW day⁻¹, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41.0 ± 9.6%, 40.5 ± 9.9% and 21.3 ± 7.4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price.     

Characterization of <Emphasis Type="Italic">kiss2</Emphasis> and <Emphasis Type="Italic">kissr2</Emphasis> genes and the regulation of kisspeptin on the HPG axis in <Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>

Reproduction allows organisms to produce offspring. Animals shift from immature juveniles into mature adults and become capable of sexual reproduction during puberty, which culminates in the first spermiation and sperm hydration or ovulation. Reproduction is closely related to the precise control of the hypothalamic–pituitary–gonadal (HPG) axis. Kisspeptin peptides are considered as the important regulator of HPG axis in mammalian. However, the current understanding of kisspeptin in flatfish is not comprehensive. In this study, we cloned and analyzed the kiss2 and kissr2 genes in Cynoglossus semilaevis. Interesting alternative splicing in the 5′-untranslated regions (UTR) of the Cskissr2 gene was found. The expression profiles of Cskiss2 and Cskissr2 showed relative high messenger RNA (mRNA) levels at the late gastrula stage during embryonic development, at total length = 40 mm during early gonadal differentiation, and in the brains and gonads of all investigated tissues. These results suggested that the kisspeptin system participated in embryogenesis and in the regulation of gonadal differentiation and development. Considering that the control and regulatory mechanisms of kisspeptin in the central reproductive axis are still unclear, we documented that the intramuscular injection of kisspeptin caused different sGnRH and cGnRH mRNA levels in a dose- and tissue-dependent manner. The mRNA expressions of FSH and LH were stimulated in the ovary and were inhibited in the testis under the kisspeptin treatments. These results provided foundations for understanding the roles of kisspeptin in the neuroendocrine system in fish. The manipulation of the kisspeptin system may provide new opportunities to control the gonadal development and even reproduction in fish.     

Growth and survival of grouper <Emphasis Type="Italic">Epinephelus</Emphasis><Emphasis Type="Italic">coioides</Emphasis> (Hamilton) larvae fed free-living nematode <Emphasis Type="Italic">Panagrellus redivivus</Emphasis> at first feeding

The free-living nematode, Panagrellus redivivus, was tested as live food for grouper Epinephelus coioides larvae during the first feeding stage. A series of experiments were conducted to determine the acceptability of the free-living nematodes in grouper larvae at first feeding, the optimum nematode density and the response of the larvae to nutritionally enriched nematode. All experiments were conducted in 200-L conical tanks filled with 150-L filtered seawater and stocked at 15 larvae L⁻¹. Duration of feeding experiments was up to day 21 (experiment 1) and 14 days (experiment 2 and 3). Brachionus plicatilis and Artemia (experiment 1) and Brachionus plicatilis alone (experiment 2 & 3) was used as the control treatment. Observations indicated that the grouper larvae readily fed on free-living nematodes as early as 3 days posthatching, the start of exogenous feeding. Optimum feeding density for the larvae was 75 nematodes ml⁻¹. The enrichment of cod liver oil or sunflower oil influenced the total lipids and n-3 highly unsaturated fatty acids of P. redivivus, which in turn influenced those of the grouper larvae, however, growth and survival of the larvae were not affected (P > 0.05). The results from this investigation showed that the nematode, P. redivivus, can be used as first live food for grouper larvae from the onset of exogenous feeding until they could feed on Artemia nauplii.     

Effect of <Emphasis Type="Italic">Cratoxylum formosum</Emphasis> on innate immune response and disease resistance against <Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> in tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

The effect of aqueous extract of Cratoxylum formosum on innate immune response and disease resistance in tilapia Oreochromis niloticus was investigated. The fish were fed diets containing 0% (control), 0.5% (diet 1), 1% (diet 2), and 1.5% (diet 3) of C. formosum aqueous extract for 30 days. At the end of the experimental period, parameters of innate immune response including phagocytosis of blood leukocytes, lysozyme activity in plasma, and respiratory bust activity were examined. Feeding the fish with diet 2 and diet 3 for 20 days enhanced phagocytic activity and for 30 days stimulated lysozyme and respiratory burst activities. Diet 1 increased the phagocytic activity at 30 days, but did not affect the other measured parameters. All parameters were not significantly changed (P > 0.05) in the control group throughout the experiment. Following 30 days of feeding, fish were infected with S. agalactiae. The cumulative mortalities of bacterial-infected tilapia that were fed diet 1, diet 2, and diet 3 were 56, 12, and 10%, respectively, compared with 85% in the control group. These results indicate that the aqueous extract of C. formosum may elevate the innate immune response and enhance disease resistance in tilapia.     

Potential of cinnamon (<Emphasis Type="Italic">Cinnamomum verum</Emphasis>) oil to control <Emphasis Type="Italic">Streptococcus iniae</Emphasis> infection in tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

In this study, four essential oils—cinnamon oil, leech lime oil, lemongrass oil, and turmeric oil—were examined for their antimicrobial activities against Streptococcus iniae, a bacterium that is pathogenic in fish, in which it causes streptococcosis. Cinnamon oil was the most potent antimicrobial agent among these oils, with a minimal inhibitory concentration (MIC) of 40 μg/ml. By using gas chromatography–mass spectrometry (GC–MS), it was found that the major components of cinnamon oil were cinnamaldehyde (90.24), limonene (2.42%), cinnamyl acetate (2.03%), linalool (1.16%), and α-terpineol (0.87%). Of these compounds, only cinnamaldehyde exhibited antimicrobial activity against S. iniae, with an MIC of 20 μg/ml. In an in vivo trial, no mortality was apparent in fish fed on fish diets supplemented with 0.4% (w/w) of cinnamon oil and with 0.1% (w/w) of oxytetracycline 5 days prior to infection with S. iniae. These results indicate that cinnamon oil had a protective effect on experimental S. iniae infection in tilapia, and thus has the potential to replace the antibiotics used to control this disease.     

Use of a <Emphasis Type="Italic">Noctiluca</Emphasis>-killing bacterium <Emphasis Type="Italic">Marinobacter salsuginis</Emphasis> strain BS2 to reduce shrimp mortality caused by <Emphasis Type="Italic">Noctiluca scintillans</Emphasis>

The ability of an algicidal bacterium Marinobacter salsuginis strain BS2, isolated from shrimp pond water, to reduce shrimp mortality was investigated under laboratory conditions. When two species of shrimp (Penaeus monodon and Litopenaeus vannamei) (body length 1.5–1.8 cm) were cultured together with the dinoflagellate Noctiluca scintillans, nearly 80 % of the shrimps died within 7 days. However, when bacterial strain BS2 was also added to the culture, N. scintillans was killed within 48 h, and shrimp survival rates on the 7th day improved from 23 to 87 % for both P. monodon and L. vannamei. The bacterium BS2 alone had no effect on shrimp condition. Under conditions of increased dissolved oxygen, the effect of using BS2 was greater, and shrimp survival rates improved even more dramatically, from 26 to 98 %. These studies provide the first evidence that the use of killing bacteria, isolated from shrimp culture water, can suppress harmful algal blooms (HABs) and thus restore the efficiency of shrimp production. The control of HABs in this way in shrimp culture farms would be a major benefit for shrimp production.     

Digestive enzymes and in-vitro digestibility of different species of phytoplankton for culture of the freshwater pearl mussel, <Emphasis Type="Italic">Hyriopsis</Emphasis> (<Emphasis Type="Italic">Hyriopsis</Emphasis>) <Emphasis Type="Italic">bialatus</Emphasis>

Hyriopsis (Hyriopsis) bialatus has been cultured during the mussel life cycle from glochidia to the adult stage with a low total survival of 6% up to 130-day-old juveniles. The main digestive enzymes (amylase and proteinases) were not detectable in one-day-old juveniles, and increased during development. The stomach, including digestive glands, was the major digestive organ for both carbohydrate and protein. The optimum conditions for amylase activity were 40°C and pH 7; for acidic proteinases they were 60°C and pH 5. Two main alkaline proteinases were found in the intestine, with optimum conditions of 30°C and pH 8 and 60°C and pH 8. To improve mussel survival by finding suitable phytoplankton species and age as food for juveniles and adults, an in-vitro digestibility test was performed on ten algal species three and seven days old using amylase and proteinases in crude enzyme extracts from different mussel life stages. Among the phytoplankton selected, the three most efficiently digested by juveniles were seven-day-old Chlorella sp.2, seven-day-old Chlorococcum sp. and seven-day-old Kirchneriella incurvata, in the ratio 1:1:3 for 30-day-old juveniles and 3:1:1 for 130-day-old juveniles. For the adult mussel, three-day-old Chlorella sp.2, seven-day-old Coccomyxa sp., and seven-day-old Monoraphidium sp., in the ratio 3:1:1, were the most digestible phytoplankton. Levels of in-vitro digestibility were related to the quality (not the concentrations) of carbohydrate and protein in the phytoplankton mixtures, and protein digestibility seemed to be the key factor determining food quality for the mussel.