Recovery of Serratia marcescens in Hemolymph of Macrobrachium rosenbergii from Experimentally Seeded Water

Seratia marcescens was used as an indicator to determine if different concentrations of bacteria in water relate to concentrations in the hemolymph of Macrobrachium rosenbergii . Prawns were exposed to high (10⁵ to 10⁶ cells/ml), low (10³ to 10⁴ cells/ml) and control (no S. marcescens ) treatments with S. marcescens inoculated in aquaria water. S. marcescens ranged from 10 to 9.9 ± 10³ cells/ml from the high concentration group; a single isolate and zero S. marcescens were recovered from the low and control groups, respectively.     

Influence of photoperiod, frequency of stripping and presence of females on sperm output in turbot, Scophthalmus maximus (L.)

Abstract. The effect of four environmental conditions was investigated upon sperm output in turbot, Scophthalmus maximus (L.), submitted to three different rhythms of stripping. Males kept under a natural light cycle and under a 6-month contracted light programme released a similar sperm output in terms of total volume of semen produced per fish during the experimental period (4·9 ± 0·9ml), mean sperm concentration (29·4 ± 2·8 × 10⁹ spermatozoa/ml) and total sperm number (163·2 ± 40·5 × 10⁹ spermatozoa). Attempts to stimulate spermiation for a second time just after the end of the natural reproduction period resulted in the release of low sperm output (total volume of semen: 1·6 ± 0·4 ml; mean sperm motility: 2 min 36s ± 0 min 47s; mean sperm concentration: 47·6 ± 10·2 × 10⁹ spermatozoa/ml; total sperm number: 84·5 ± 25·3 × 10⁹ spermatozoa). Stripping frequency had no effect on total volume of semen, mean sperm motility and total sperm number. Monthly collection did not modify sperm samples in relation to stripping rank. However, decreasing volume, motility and sperm concentration were observed when males were stripped fortnightly and weekly. During the natural spawning period, the presence of females in the tank enhanced mean sperm motility (from 3 min 27s + 0 min 52s to 6 min 38s ± 1 min 38s).     

Enrichment of Rotifers Brachionus plicatilis with Eicosapentaenoic Acid and Docosahexaenoic Acid Produced by Bacteria

Abstract— Two bacterial strains, rich in either eicosapentaenoic acid [EPA, 20:5(n-3)] ( Shewanella gel-idimarina ACAM 456) or docosahexaenoic acid [DHA, 22:6(n-3)] ( Colwellia psychroeryrhrus ACAM 605) were tested for their ability to enrich rotifers Erachionus plicatilis in these polyunsaturated fatty acids. Rotifers were exposed for 24 h to each bacterial strain and to a mixture of the two strains. They were then harvested and their fatty acid compositions were analysed and compared to those of rotifers that had been either starved or fed yeast Saccharomyces cerevisiae or microalgae Tetraselmis suecica in 2-L glass flasks. Exposure to 1.4 × 10⁹ cells/ml of the EPA-producing bacterium only resulted in rotifer EPA levels increasing from 0.1% to 1.2% of total dry weight (%dw). Similarly, following exposure to 1.0 × 10⁹ cells/mL of the DHA-producing bacterium only, rotifer DHA levels increased from below detection to 0.1% dw. When exposed to a mixture of the two bacterial strains, containing 7.0 × 10⁸ celldml of the EPA producer and 5.0 × 10⁸ cells/mL of the DHA producer, the rotifers'final EPA and DHA levels were 0.5% dw and 0.3% dw respectively. Although feeding strategies need refining, these results show, for the first time, that rotifers can be enriched with DHA from bacteria, and that rotifers can be enriched simultaneously with both DHA and EPA from different bacterial strains.     

Fertilization efficiency of cryopreserved sperm from striped catfish, Pangasius hypophthalmus (Sauvage)

The fertilization efficiency of cryopreserved sperm was compared with fresh sperm from striped catfish, Pangasius hypophthalmus . Of the two sets of experiments carried out, the first compared four sperm doses using fresh sperm and fresh eggs. The second experiment compared six concentrations of cryopreserved sperm ranging from 6.94 × 10⁷ to 6.94 × 10¹⁰ to fertilize 100 eggs per batch. Fertilization, hatch and survival rates were compared between cryopreserved and fresh sperm. The highest fertilization rate (53.75±1.62%) was achieved with a sperm dose of 6.94 × 10⁸. Increasing the sperm dose to 3.47 × 10⁹ did not increase the fertilization rate, indicating that the optimum sperm:egg ratio lies between 6.94 × 10⁶ and 3.47 × 10⁷ sperm per egg. Both highest (6.94 × 10¹⁰) and the lowest (6.94 × 10⁷) sperm doses resulted in lower fertilization rates (2.04% and 16.90% respectively). No significant differences were found among four fresh sperm doses compared. Mean hatch and survival rates resulting from fresh and cryopreserved sperm were similar. The experiment shows that while only 1.89 × 10⁶ fresh spermatozoa was required to fertilize a fresh egg, 6.94 × 10⁶ (or 3.67 times more) cryopreserved sperm was required to achieve the same level of fertilization. This provides important information for making decision to cryopreserve sperm for commercial and/or conservation purposes.     

Continuous exposure to infectious pancreatic necrosis virus during early life stages of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss (Walbaum), were exposed continuously to infectious pancreatic necrosis virus (IPNV) at 0, 10¹, 10³ or 10⁵ plaque forming units (pfu) L⁻¹ of water to estimate the effects of chronic IPNV exposure on early life stages. Fish density averaged 35 fish L⁻¹ (low density) or 140 fish L⁻¹ (high density), and the tank flow rate was 250 mL⁻¹ min. Virus exposure began at 6 days before hatch and continued until fish were 44 days old. Cumulative per cent mortality, analysis of survival and hazard functions, and discrete-time event analysis were used to explore the patterns of survival and mortality. In eggs and fish exposed to IPNV, mortality significantly greater than in the 0 pfu L⁻¹ exposure did not occur until IPNV concentration was 10⁵ pfu L⁻¹ at low fish density and 10³ pfu IPNV L⁻¹ at high fish density. These results suggest that in the natural aquatic environment, where rainbow trout densities are likely to be considerably lower than in this study, mortality resulting from infection with IPNV will very likely not occur when ambient concentrations of virus are ≤10³ pfu IPNV L⁻¹. In aquaculture rearing units, trout density is likely to be as high or higher than the densities used in this study. Therefore, continuous inputs of virus at concentrations greater than 10¹ pfu L⁻¹ may result in IPN epidemics in aquaculture facilities.     

The enrichment and retention of ascorbic acid in rotifers fed microalgal diets

The enrichment and retention of ascorbic acid (AA) was investigated in rotifers Brachionus plicatilis fed on microalgae ( Nannochloropsis oculata and Isochrysis sp. (T.ISO)) and baker's yeast Saccharomyces cerevisiae . The concentrations of AA of the rotifer diets used in the study differed significantly: 4200 μg g⁻¹ of dry weight in Isochrysis sp. (T.ISO), 2600 μg g⁻¹ in N. oculata and only 77 μg g⁻¹ in S. cerevisiae . Rotifers contained 620 μg AA g⁻¹ prior to the experimental feeding. When subsequently fed for 3 h on microalgae at a ration of 0.13 mg dry microalgae per 10⁶ rotifers rapidly and efficiently increased their content of AA: Isochrysis sp.-fed rotifers contained 1600 μg AA g⁻¹ and N. oculata -fed rotifers contained 1100 μg AA g⁻¹. Concentrations were boosted by a further feeding of a second ration of algae at three times the initial feeding ration; 21 h later, Isochrysis sp.-fed rotifers contained 2500 μg AA g⁻¹ and N. oculata -fed rotifers contained 1700 μg AA g⁻¹. This represented a 180% and 310% increase in the pre-feeding vitamin concentrations in Isochrysis sp. and N. oculata -fed rotifers, respectively. There were no significant changes in AA concentration in rotifers fed a similar ration of yeast throughout the feeding period (520-620 μg AA g⁻¹). Rotifers retained AA during a subsequent 24 h non-feeding period, with no significant changes in the concentrations in any of the rotifer groups. The production of rotifers rich in AA may be particularly valuable for the culture of fish larvae that have a high requirement for the vitamin.     

Studies on the pathogenesis of streptococcal infection in cultured yellowtails Seriola spp.: effect of the cell free culture on experimental streptococcal infection

Abstract. A previous paper has revealed that experimental streptococcal infection was associated with a rapid growth of the intestinal bacteria, suggesting association of the condition with an exotoxin. The present study was undertaken to see the effect of the exotoxin on the streptococcal infection by administering cell free culture before the bacterial challenge. Cell free culture of Streptococcus sp. strain YT-3 was inoculated intramuscularly at a dosage of 0–5 ml per 100 g body weight 1 h prior to bacterial challenge. Three fish were killed at intervals after challenge for viable counting of bacteria in the viscera and blood. Intramuscular inoculation with either low virulent or virulent bacteria alone at dosages of 10⁶ to 10⁷ cells did not produce the disease in the fish, with almost complete clearance of the bacteria from the viscera and blood within 120 h. When the exotoxin was inoculated intramuscularly prior to the bacterial challenge, however, either low virulent or virulent bacteria at 10⁶ to 10⁷ cells could produce fatal infection with prominent streptococcal clinical signs.     

The effect of dietary phosphatidylcholine and its constituent fatty acids on microdiet ingestion and fatty acid absorption rate in gilthead sea bream, Sparus auratus, larvae

This study tested the effect of the level of dietary phosphatidylcholine (PC) and its constituent medium-chain fatty acids on microdiet ingestion (μg diet larva⁻¹ h⁻¹) and the absorption rate of the free fatty acid [¹⁴C]16:0 (pmole larva⁻¹ h⁻¹) in 15, 20, 21, 25, 26, 30 and 31-day-old gilthead sea bream, Sparus auratus L., larvae. Fish were fed four microdiets (A, B, C and D): microdiet A contained no phospholipid (PL), while microdiet B included 10 g kg⁻¹ Artemia nauplii PL (3.7 g kg⁻¹ PC). Microdiets C and D contained 10 g kg⁻¹ purified saturated PC dimyristoyl (C_14:0) and polyunsaturated PC dilinoleoyl (C_{18:2[cis]−9,12}), respectively.
Larvae from one or both of the PC microdiets demonstrated significantly higher ( P < 0.05) ingestion rates (μg diet larva⁻¹ h⁻¹) than the non-PL microdiet control in 15, 21, 22, 25 and 26-day-old larvae and the Artemia PL microdiet in 15, 22 and 26-day-old larvae. However, microdiet ingestion and fatty acid absorption rate appeared to be independent of the associated medium carbon chain saturated or polyunsaturated fatty acid moiety of the PC diets. Apparent absorption, as measured by the retention of radio-labelled [¹⁴C]16:0 following 8 h of non-labelled microdiet feeding, was possibly related to feeding.     

Water Quality and Microbial Dynamics in Shrimp Ponds Receiving Bagasse-Based Feed

Pond water quality and associated microbial biomass were studied in relation to the type of feed applied during the culture of the marine shrimp Penaeus vannamei . The feeds tested included conventional feedlot manure as well as two feeds based on bagasse, a sugarcane waste product. Physical and chemical parameters were studied during a 100 day trial in 200 m² earthen ponds. Both bagasse-based feeds supported a significantly larger microbial community as measured by specific biomass numbers ( P < 0.01), ATP content ( P < 0.001) and amount of the particulate organic matter present on pond bottoms ( P < 0.025). For both bagasse-based treatments, the estimated bacterial cell number in the flocculent layer was 3.11 ± 10¹²/m², compared to the much lower cell number of 7.53 ± 10¹⁰/m² for control ponds. Harvest data suggest that bagasse forms a potential base for feeds when applied to extensive shrimp cultures.     

Preliminary investigations on husbandry, nutrition and growth of glass eels and elvers, Anguilla anguilla L.

Abstract. Some effects of husbandry and feeding on growth of glass eels and elvers of the European eel. Anguilla anguilla L., were investigated. In the first 7 months after arrival the recorded mortality amounted to 34% of the initial population. due to escapes and possibly cannibalism the observed survival after 7 months was only 21% of the initial population.
Mean growth rate of the whole population was 7.1 g day⁻¹ kg^−0.8. Growth proved to be highly variable. Growth rate was clearly affected by feeding level but not by initial stocking density.
Maximum growth rate was approximately 8g day, ¹ kg^−0.8 with feed conversions of 1.7–2.0. An optimal feeding level of 7.5g day¹ kg^−0.8 resulted in feed conversions of 1.3–1.6.     

Effect of water temperature on the growth performance and digestive enzyme activities of Chinese longsnout catfish (Leiocassis longirostris Günther)

The present study was carried out to investigate the influence of water temperature on the growth performance and digestive enzyme (pepsin, trypsin and lipase) activities of Chinese longsnout catfish. Triplicate groups of Chinese longsnout catfish (35.6±0.48 g, mean±SE) were reared at different water temperatures (20, 24, 28 and 32 °C). The feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) were significantly affected by water temperatures and regression relationships between water temperature and FI, SGR as well as FER were expressed as FR=−0.016 T ²+0.91 T −10.88 ( n =12, R ²=0.8752), SGR=−0.026 T ²+1.39 T −17.29 ( n =12, R ²=0.7599) and FER=−0.013 T ²+0.70 T −8.43 ( n =12, R ²=0.7272). Based on these, the optimum temperatures for FR, SGR and FER were 27.66, 26.69 and 26.44 °C respectively. The specific activities of digestive enzymes at 24 or 28 °C were significantly higher than that at 20 or 32 °C. In addition, there was a significant linear regression between FR or SGR and specific activities of pepsin and lipase, which indicated that pepsin and lipase played important roles in regulating growth through nutrient digestion in Chinese longsnout catfish.     

Cage culture of the Pacific white shrimp Litopenaeus vannamei (Boone, 1931) at different stocking densities in a shallow eutrophic lake

Postlarvae of Litopenaeus vannamei were acclimated and stocked in lake-based cages at the following stocking densities: 10, 20, 30 and 40 shrimp m⁻². Another set of shrimp was stocked in concrete tanks as reference samples at 30 shrimp m⁻². Significant differences were observed among stocking densities throughout the 95-day culture. The final weight at harvest decreased with increasing stocking density: mean weights of 23.3, 15.8, 13.0, 10.9 and 14.6 g for the 10, 20, 30, 40 shrimp m⁻² and reference tanks were observed respectively. There were no significant differences in survival throughout the culture period, ranging between 69% and 77%. Daily growth rates (range: 0.11–0.24 g day⁻¹) and specific growth rates (range: 3.54–4.34%) also differed significantly among stocking densities, both increasing with decreasing stocking density. The feed conversion ratio in the cages did not differ among the stocking densities, ranging from 1.53 to 1.65. The relationship between stocking density and mean individual weight at harvest followed the equation y =81.06 x ^−0.54 ( R ²=0.938) and that of stocking density and production (in g m⁻²) is y =58.01 x ^−0.46 ( R ²=0.834).     

Growth of Crawfish Procambarus clarkii as a Function of Density and Food Resources

This study utilized enclosures (cylindrical, 5-mm wire mesh, O.5 m² bottom surface area) placed over rice-forage substrates in experimental crawfish ponds to contain crawfish under typical pond culture conditions. Juvenile Procambarus clarkii were stocked at six densities (2, 4, 6, 10, 14, and 18 crawfish/m²) for 12-wk growth trials in October and again in February. Crawfish relied solely on the detrital food system for their nutritional needs. Supplemental feed was supplied to crawfish in additional enclosures at two of the test densities (2 and 10 crawfish/m²). The commercially formulated feed (25% crude protein) was fed (2.02 g dry feed/m²) 3 d/wk (Monday, Wednesday, Friday). All treatments were replicated with six enclosures. Crawfish growth was inversely correlated to culture density. Mean final weights for crawfish feeding from the detrital-system only were 15.3, 13.8, 11.2, 7.9, 7.2, and 5.8 g for crawfish densities of 2, 4, 6, 8, 10, 14, and 18/m², respectively. Mean final weights for crawfish receiving supplemental feed were 20.7 and 12.4 g for densities of 2 and 10 crawfish/m². When compared with density as a factor influencing growth, feed influenced growth less than density abatement. Supplemental feeding improved crawfish growth in detrital systems an average of 46%, while decreasing initial density improved growth an average of 80.5%.     

Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms

Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus , caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10⁷ copies μg⁻¹ DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10⁶–10⁸ copies of CyHV-2 μg⁻¹ DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22–10 °C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (±SE) of 7.3 ± 11 to 394 ± 55 μg⁻¹ DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease.     

The infectivity by different routes of exposure and shedding rates of Aeromonas salmonicida subsp. salmonicida in Atlantic salmon, Salmo salar L., held in sea water

Abstract. The infectivity of the bacterial fish pathogen Aeromonas salmonicida subsp. salmonicida to Atlantic salmon, Salmo salar L., in sea water was investigated and found to be similar to that reported for fresh water. The minimal infective dose in short duration bath exposures (1–3 days) was 10⁴ colony-forming units (cfu) per ml, while prolonged exposure for three weeks, but not for 1 week, produced infection with 10² cfu/ml. Intragastric intubation of A. salmonicida established infection with doses of >10⁵ cfu. Release of bacteria from dead or morbid infected fish was monitored and found to be in the order of 10⁵–10⁸ cfu/fish/h. These results emphasize the importance of removing dead fish from farm sites.     

The Intensive Culture of the Penaeid Shrimp Penaeus vannamei Boone in a Recirculating Raceway System

Recirculating raceway systems were examined for their potential as a method for the intensive culture of the marine shrimp Penaeus vannamei Boone. The systems consisted of fiberglass raceways 38 m³ (13.7 m ± 2.4 m ± 1.16 m) and 28 m³ (13.7 m ± 2.4 m ± 0.85 m) each equipped with a vertical screen biofilter, foam fractionators and an ultraviolet ozone generator. All of the systems were enclosed in a commercial greenhouse. Four preliminary growout experiments and two growout experiments with stocking densities of 970 shrimp/m³ and 2,132 shrimp/m³ were completed.
Temperature, pH and salinity remained constant throughout the experiments. Unionized ammonia levels remained below 0.2 mg/L. Nitrite levels ranged from 0.1 to 1.0 mg/L. The 2,132/m² stocking density resulted in 48% survival, food conversion ratio (FCR) of 1.8 and an average size of 10.8 g. The 970/m³ stocking density resulted in 82% survival, FCR of 2, and an average size of 14 g. Production was 11.4 kg/m³ (114 tons/ha) and 11.0 kg/m³ (110 tons/ha) for the high and low stocking densities, respectively.     

The susceptibility of salmonid fish to an atypical strain of Aeromonas salmonicida that infects goldfish, Carassius auratus (L.), in Australia

Abstract. An enzootic, Australian, atypical strain of Aeromonas salmonicida isolated from diseased goldfish, Carassius auratus (L.), was inoculated into Atlantic salmon, Salmo salar L., brown trout, S. trutta L., rainbow trout, S. gairdneri Richardson, and brook trout, Salvelinus fontinalis (Mitchill), fingerlings by intraperitoneal injection (i.p.) and by bath challenge, the latter with and without prior abrasion of skin. The 10-day LD₅₀ (i.p.) was estimated to be 7·4 × 10^-3 colony forming units (cfu) for Atlantic salmon, 3·0 × 10^-2 cfu for brown trout, 3·7 × 10² cfu for brook trout and 6·4 × 10³ cfu for rainbow trout. Brown, rainbow and brook trout succumbed to bath challenges with between 10⁵–10⁶ cfu/ml, developing ulcers of the skin and septicaemia. The organism was trasmitted from inoculated fish to five of 195 within-tank control fish via water and established a carrier state in one of 14 Atlantic salmon. It was concluded that the organism poses a significant threat to the salmonid farming industry and wild salmonid fisheries in Australia.     

Effect of antibiotic treatment on the growth and survival of juvenile northern Chilean scallop, Argopecten purpuratus Lamarck (1819), and associated microflora in experimental cultures

Juvenile northern Chilean scallops of 937±55 μm shell height were exposed to five different concentrations of chloramphenicol (CHL) (5, 10, 25, 50 and 100 μg mL⁻¹), plus a control without antibiotics. To determine the effect of CHL on the accompanying microflora, the number of colony-forming units (CFU) that grew on TGE culture medium was counted in the seawater of containers with juveniles, and in containers with microalgae used as food. Both were exposed to the same concentrations of CHL. The growth rates of juveniles treated with CHL and the control without antibiotic showed highly significant differences ( P =0.0001). The growth rate was inversely proportional to the CHL concentration. The control sample presented the highest growth rate (84.4±14.3 μm day⁻¹), followed by the sample treated with 5 μg mL⁻¹ (64.2±14.3 μm day⁻¹). The survival in the control and in the treated samples with 5–50 μg mL⁻¹ was rather high, with a mean value of 95%. Only the sample treated with 100 μg mL⁻¹ had a low survival (36.7%). The CFU count was larger in the containers with juveniles, when compared with the ones with food. The CFU count tended to decrease with increasing CHL concentration in the juveniles.     

Patterns of stable carbon and nitrogen isotopes in the baleen of common minke whale Balaenoptera acutorostrata from the western North Pacific

ABSTRACT:   Stable carbon and nitrogen isotope ratios were determined in the baleen plates of 17 common minke whales Balaenoptera acutorostrata from the north-western Pacific Ocean off Japan, as well as prey species (krill Euphausia pacifica , Japanese anchovy Engraulis japonicus and Pacific saury Cololabis saira ) collected in the stomach contents, to investigate the trophic relationship between the minke whales and their prey. A few δ¹⁵N-depleted peaks occurred along the length of baleen plates for 10 males irrespective of stomach content (anchovies and sauries). Similar δ¹⁵N-depleted peaks were also found for one female and two immature individuals. It was likely that these δ¹⁵N-depleted peaks formed in early summer. The stable nitrogen isotope ratio (δ¹⁵N) values in Pacific saury (9.3 ± 1.4‰) did not differ significantly from that in Japanese anchovy (8.8 ± 0.9‰). In contrast, δ¹⁵N in krill (7.2 ± 0.5‰ in July and 8.0 ± 0.2‰ in September) were significantly lower than in the Pacific saury. Thus, these peaks may reflect the dietary change from krill to fishes in the feeding migration of the whales. Growth rate of the baleen plate was estimated to be 129 mm/y, and it appeared that a dietary record of about 1.4 years remained in the baleen plate. For two immature whales, the maximum value of δ¹⁵N occurred at the tip of baleen. This δ¹⁵N enrichment may possibly be useful for discriminating weanlings and older whales.     

Effects of Water Temperature, Density, and Sex Ratio on the Spawning Rate of Red Claw Crayfish Cherax quadricarinatus (von Martens)

Effects of water temperature on red claw crayfish Cherax quadricarinatus spawning rates was evaluated for 3 mo at 22, 26 and 30 ± 1 C. A 20/m² stocking density and a 1:2 M: F sex ratio was used. Spawning rate increased as temperature increased, with 30 C providing the highest spawning rate ( P < 0.05). Effects of stocking density and sex ratio on the reproduction of red claw crayfish were also evaluated at densities of 10.15 and 20/m², with a sex ratio of 1:1, and at three sex ratios (1:1. 1:3 and 1:5 M: F) at a density 10/m². Water temperature was maintained at 28 ± 1 C and photoperiod was maintained at 14L: 10D for 6 mo during the density and sex ratio experiments. Spawning rates were not significantly different ( P > 0.05) within the three stocking densities at a sex ratio of 1:1, nor were they different ( P > 0.05) for the three sex ratios at a density of 10/m². Results revenled that densities as high as 20/m² and sex ratios of 1 male to 5 females can be used to increase spawning per tank. Also, high temperature (28 ± 1 C) and long day lengths (14L: 10D) can be utilized to stimulate peak spawning rates, but high spawning rates cannot be maintained for more than about 3 mo.