Effect of different live food on survival and growth of first feeding barber goby,Elacatinus figaro (Sazima,Moura & Rosa 1997) larvae

One of the major challenges in marine fish culture is how to provide live food of adequate size and nutritional quality for first‐feeding larvae. Commonly used live food organisms, rotifers and brine shrimp, may not always be the best option. To determine the suitability of different zooplankton in the larviculture of Elacatinus figaro, three diets were tested: RE – rotifers Brachionus sp. (10 ind mL^?1)+ciliate Euplotes sp. (10 ind mL^?1), enriched with fatty acids; RC – enriched rotifers (10 ind mL^?1)+wild copepod nauplii (10 ind mL^?1); and R – enriched rotifers (20 ind mL^?1). Survival rates were estimated 10 days after hatch (DAH) for the three test groups, and growth rates were evaluated for RE and R at 10 and 20 DAH. Although survival rate was numerically higher for the RC diet (41.1±14.2%), no significant difference was detected between groups fed RE (20.5±18.1%), RC or R (32.1±16.5%). At 10 DAH, the growth rate was significantly higher in RC (5.7±0.6 mm) than in R (4.6±0.5 mm), a trend that was also observed at 20 DAH for RC (8.6±0.5 mm) and R (5.8±0.7 mm) (P<0.05). E. figaro larvae fed on ciliates did not show satisfactory results, whereas feeding copepod nauplii enhanced growth.     

Sperm quality of Brazilian flounder Paralichthys orbignyanus throughout the reproductive season

The aim of this study was to evaluate the sperm quality of Brazilian flounder Paralichthys orbignyanus throughout its reproductive season. Sperm was collected at the beginning, middle and end of the breeding period. Spermatozoa density was maximum at the beginning (12.7 ± 0.92 × 10⁹ cells mL^?1) and at the end (11.8 ± 0.39 × 10⁹ cells mL^?1) of the breeding season (P<0.05). Sperm production and the percentage of spermatozoa moving fast forward increased significantly towards the end of the breeding season (P<0.05). The mean duration of progressive motility of spermatozoa was around 10 min. No difference was observed during the reproductive season in the percentage of motile cells, pH, osmolality and K⁺, Cl^? and Mg²⁺ concentrations in seminal plasma. The concentration of Na⁺ increased throughout the breeding season, reaching 174.62 ± 12.68 mmol L^?1 at the end (P<0.05). There was a decline in the concentration of Ca²⁺ (12.31 ± 3.08 mmol L^?1) in the middle of the breeding season, which coincided with the shortest motility duration of spermatozoa. The information reported in this study should help to improve management and optimize the development of protocols for short‐term storage and cryopreservation of Brazilian flounder semen.     

Influence of stocking density and temperature on early development and survival of the purple sea urchin,Strongylocentrotus purpuratus (Stimpson, 1857)

We evaluated the effect of four densities (940, 1880, 3760, 7520 eggs cm^?2 and 0.5, 1, 2, 4 ind mL^?1 of embryos and larvae, respectively) and four temperatures (8, 11, 14, 17°C) on early growth and survival of the sea urchin Strongylocentrotus purpuratus. Prism‐stage length was significantly greater in embryos initially held at 940 and 1880 eggs cm^?2 than in those held at 3760 and 7520 eggs cm^?2. Larvae grew significantly faster and had significantly greater survival when reared at 0.5 or 1 ind mL^?1 than when held at 2 or 4 ind mL^?1. Embryos had greater survival at 11 and 14°C than at 8 and 17°C, whereas embryo length was significantly smaller at 8°C than at 11, 14 or 17°C. Larvae grew significantly slower at 8°C than at 11, 14 or 17°C, whereas survival was significantly reduced at 8 and 17°C compared with 11 and 14°C. Per cent survival from prism to metamorphic competency in the best treatments was 48.9 ± 2.2% and 50.0 ± 3.6% (mean ± SE) for the 1 ind mL^?1 and 11°C treatments, respectively. On the basis of these results, for rearing of S. purpuratus under static conditions, we recommend that fertilized eggs and larvae be held at ≤1880 eggs cm^?2 and ≤1 ind mL^?1, respectively, and at 11–14°C.     

Development of an enzymatic method for individual separation of banded coral shrimp (Stenopus hispidus) embryos

The outer coat of embryo of Stenopus hispidus may affect cryoprotectant penetration and post‐thaw procedures during cryopreservation studies. The mechanical method for separating embryos of S. hispidus was exhausting and time consuming. The aim of the present study was to compare the effect of different enzymatic isolation techniques to achieve the individual separation of embryos. Embryo clusters were exposed to three enzymes (trypsin, collagenase and hyaluronidase) at different concentrations (0.2–1.6 mg mL^?1) for different treatment times (5–20 min). Morphological appearance and hatching competence of the embryos separated mechanically and enzymatically were assessed. The optimum conditions for the three enzymes were 10 min 0.4 mg mL^?1 trypsin, 10 min 0.8 mg mL^?1 collagenase and 10 min 0.8 mg mL^?1 hyalruonidase and embryo hatching percentages were 43.8 ± 4.2%, 39.9 ± 4.9% and 42.2 ± 3.5% respectively. The enzymatic isolation technique does not affect either morphological appearance or hatching competence of the separated embryos. The method developed in this study will be helpful for future cryopreservation studies on S. hispidus embryos.     

An intensive hatchery rearing protocol for larvae of the bullseye puffer,Sphoeroides annulatus (Jenyns)

In this study bullseye puffer, Sphoeroides annulatus larvae were reared from hatching through to 1 or 2 months after weaning on an experimental scale in three replicate 600 L tanks and on three occasions during the spawning season (nine tanks in total). The rearing protocol used was green water (Nannochloropsis oculata and Isochrysis sp.) 100 000 cells mL^?1 from 0 to 11 days after hatch (DAH), 5–10 rotifers, Brachionus rotundiformis mL^?1 from 2 to 26 DAH, 0.1–1 Artemia mL^?1 from 21 to 34 DAH and weaning from 29 to 34 DAH. Survival to a month after weaning was 1%, a total of 3153 juveniles were produced with an average wet weight of 0.29 ± 0.07 g and a length of 27.5 ± 0.82 mm.     

Chemical surface disinfection of eggs of Baltic cod,Gadus morhua L.

The effect of two disinfectants on eggs and larvae of Baltic cod, Gadus morhua, was investigated. The eggs were disinfected for 10 min using various concentrations of either glutaraldehyde (100, 200, 400, 600 and 800 mg L^?1) or iodophor (10, 50, 100 and 150 mg L^?1), 1–4‐days post‐fertilization. Bactericidal effect of disinfection, survival to hatching, hatching success and larval abnormalities were assessed. Larval survival was recorded at 5‐, 10‐ and 15‐days post‐hatch (dph). Although Baltic cod eggs have an unusually thin chorion, they could tolerate surface disinfection. A reduction in bacterial growth was observed with increased concentrations of disinfectant (3.0 × 10⁷–1.6 × 10¹ CFU mL^?1). Abnormalities in newly hatched larvae were not related to disinfection. Survival of the yolk sac larvae was significantly better for eggs treated with 400 mg L^?1 glutaraldehyde for 10 min at 10 and 15 dph. Effective disinfection was also recorded using 100 mg L^?1 Actomar K30. Egg batch effect rather than initial bacterial concentration, disinfectant type or incubation method determined the survival of the eggs to hatching and survival of larvae. Because of the carcinogenic effect of glutaraldehyde, iodophor is recommended for routine disinfection of cod eggs.     

Occurrence of Listonella anguillarum in seed production environments of Japanese flounder Paralichthys olivaceus (Temminck et Schlegel)

The present study was undertaken to investigate the distribution of Listonella anguillarum in the rearing water, fish and diets (rotifers) of Japanese flounder (Paralichthys olivaceus). A total of 793 isolates were obtained from the seed production environment of Japanese flounder and 175 out of them were identified as L. anguillarum by biochemical characterization, polymerase chain reaction (PCR) detection for VAH1 haemolysin gene and phylogenetic analysis of 16S ribosomal deoxyribonucleic acids (rDNA) sequences. These results strongly suggested that L. anguillarum is rapidly and accurately identified by the combination of incubation on thiosulphate–citrate–bile salt–sucrose agar at 35°C overnight and PCR detection for the VAH1 haemolysin gene. All flounder specimens and all rotifer samples harboured L. anguillarum at high densities of 6.9 × 10³–6.3 × 10⁵ colony forming units (CFU) g^?1 and 1.5 × 10⁴–2.3 × 10⁶ CFU g^?1, respectively, while as low as 5.0 × 10⁰–2.0 × 10¹ CFU mL^?1 of L. aguillarum were detected in only two of 11 seawater samples, even though no vibriosis occurred in larval and juvenile flounder of tanks. This fact strongly suggests that L. anguillarum is an inhabitant in the seed production environments of Japanese flounder.     

Isolation and culture of the marine rotifer, Colurella dicentra (Gosse, 1887), from a Mississippi Gulf Coast estuary

Few marine rotifer species (e.g. Encentrum linheii and Synchaeta cecilia) have been cultured successfully besides Brachionus plicatilis and B. rotundiformis, commonly used to rear larvae of many marine fish species. The development of culture techniques for marine rotifers smaller in size than the Brachionus species may be useful for rearing fish species for which the currently used prey are too large. We evaluated the possibility of culturing Colurella dicentra isolated from a Mississippi Gulf Coast estuary. An experiment was conducted to determine the effects of salinity (10–35 g L^?1) on its population growth rate. Rotifers were fed Nannochloropsis oculata at a density of 100 000 cells mL^?1 for 15 days. Colurella dicentra survived in water with a salinity of 10–47 g L^?1. Densities of up to 300 rotifers mL^?1 were sometimes attained in cultures. Salinity influenced C. dicentra production (P<0.001). The mean rotifer numbers at 10 g L^?1 (22 840±2604 SD), 15 g L^?1 (25 980±7071 SD) and 20 g L^?1 (19 780±1029 SD) at the end of the experiment were similar (P>0.05), but were higher (P=0.05) than numbers at 25 g L^?1 (4240±1783), 30 g L^?1 (1300±264 SD) and 35 g L^?1 (100±101 SD). The population growth rate (r) of the rotifers was the highest at 15 g L^?1 (0.37–0.42 day^?1), and the lowest at 35 g L^?1 (?0.33–0.06 day^?1). This is the first report of C. dicentra in the estuarine waters of the Gulf of Mexico, and also the first time it has been cultured successfully.     

Stocking density evaluation on Catarina scallop (Argopecten ventricosus,Sowerby II, 1842) larvae to improve hatchery production

The Catarina scallop Argopecten ventricosus is a highly valued resource. Although its hatchery spat production has already been reported, the effects of initial larval stocking density have never been reported for production purposes. This study evaluates A. ventricosus growth and survival in triplicate using three stocking densities: low (LD; 2 larvae mL^?1), medium (MD; 4 larvae mL^?1), and high (HD; 6 larvae mL^?1). Three-day old larvae were reared in 18-L plastic carboy at 25.6?±?0.5 °C and fed with a microalgal blend of Isochrysis galbana and Chaetoceros calcitrans (1:1 cell number ratio) for 7 days, equivalent to 10 post-fertilization days (PFD). Higher specific growth rate was recorded at LD (15.8?±?0.2%) after 8 PFD of culture compared to MD (1.6?±?0.5%) and HD (4.1?±?1.8%) densities. The least time required for 60% of the larvae to reach the pediveliger stage was recorded at LD condition (10 PFD). Higher survival was recorded at HD (58.8?±?3.1%) at 8 PFD compared to MD (53.5?±?3.1%) and LD (43.9?±?3.0%). After 8 PFD, stocking density was highly related to larval growth and survival. To increase production and growth, and reduce the time required to reach pediveliger stage, stocking density should start with 6 larvae mL^?1 and be reduced to 2 larvae mL^?1 at 7 PFD.

     

Biological characteristics of the improved extensive shrimp system in the Mekong delta of Vietnam

Low and unstable shrimp yields of the improved extensive shrimp system has been a tremendous obstacle for economic development in the coastal areas of Southern Vietnam. To investigate the biological characteristics of this system, ponds in the coastal Cai Nuoc district, Mekong delta of Vietnam, were monitored. Results showed that the system was not optimal for shrimp. While chlorophyll a (chl a) (1.51–37.2 μg L^?1), phytoplankton density (6333–974 444 cells L^?1) and zooplankton density (7.1–517.2 ind L^?1) were abundant and comparable to shrimp farms elsewhere, zoobenthic community was very poor (7–1971 ind m^?2). Toxin‐producing cyanobacteria (Oscillatoria limosa, Oscillatoria formosa, Anabaena sp. and Phormidium tenue) were found. Total bacteria and Vibrios were present in large numbers (respectively 1.04 × 10⁵ and 6.64 × 10² CFU mL^?1 in pond water, 6.33 × 10⁵ and 9.47 × 10³ CFU g^?1 in sediment). Presence of toxin‐producing organisms, poor zoobenthic community and abundance of Vibrios all can enhance shrimp susceptibility to diseases. The following measures are recommended to improve the situation: (1) complete testing of seeds for pathogens, (2) not to incorporate fish into shrimp ponds and (3) applying no‐culture breaks and pathogen‐killing chemicals.     

The larval rearing of the marine ornamental crab, Mithraculus forceps (A. Milne Edwards, 1875) (Decapoda: Brachyura: Majidae)

The goal of this study is to develop a larviculture protocol for Mithraculus forceps, a popular marine aquarium species. Different temperatures (25±0.5°C and 28±0.5°C), stocking densities (10, 20, 40 and 80 larvae L^?1), prey densities (newly hatched Artemia of 1, 4, 7 and 12 nauplii mL^?1) and metamorphosis to crab conditions (Systems A and B) were tested. The best survivorship and faster development were obtained when the larvae were reared at a density of 40 larvae L^?1 for 7 days post hatching (DPH) in System A, at 28°C and fed with 7 mL^?1 of newly hatched Artemia nauplii. After 7 DPH all the megalopa were moved to System B and the same temperature and prey density were maintained. At the end of the experiment, 12 DPH, survivorship of 74.1±4.8% was obtained.     

The effect of different rotifer feeding regimes on the growth and survival of yellowtail kingfish Seriola lalandi (Valenciennes, 1833) larvae

First feeding success is critical to larval marine finfish and optimization of live feed densities is important for larval performance and the economics of commercial hatchery production. This study investigated various rotifer feeding regimes on the prey consumption, growth and survival of yellowtail kingfish Seriola lalandi larvae over the first 12 days post hatch (dph). The common practice of maintaining high densities of rotifers (10–30 ind. mL^?1) in the rearing tank was compared to a low density feeding technique, where 5–8 ind. mL^?1 of rotifers were offered. A ‘hybrid’ feeding regime offered rotifers at the high density treatment until 5 dph and the lower feeding densities thereafter. There was no significant difference in larval survival (hybrid: 28.9 ± 7%, low density: 17.3 ± 5% and high density: 17.2 ± 9%) or growth (hybrid: 6.12 ± 0.18 mm, low density: 6.03 ± 0.10 mm and high density: 6.11 ± 0.23 mm) between treatments. Rotifer ingestion was independent of rotifer density throughout the trial and increased with larval age, with larvae at 4 dph ingesting 22 ± 1.5 rotifers larvae^?1 h^?1 and by 11 dph ingesting 59 ± 1.6 rotifers larvae^?1 h^?1. These data demonstrate that from first feeding, yellowtail kingfish larvae are efficient at capturing prey at the densities presented here and consequently significant savings in rotifer production costs as well as other potential benefits such as facilitation of early weaning and improved rotifer nutritional value may be obtained by utilizing lower density rotifer feeding regimes.     

Evaluation of selected metal nanoparticles on hatching and survival of larvae and fry of Indian major carp,rohu (Labeo rohita)

Different laboratory synthesized metal nanoparticles viz. Copper oxide (CuO), Zinc oxide (ZnO) and silver doped titanium dioxide (Ag‐TiO₂) were studied for their effect on hatching and survival of larvae and fry of Indian major carp, rohu, Labeo rohita both in direct application in tank water & coated onto tanks. Among these nanoparticles, CuO and ZnO nanoparticles exhibited highest percentage of hatching in both direct addition (78.0 ± 3.1% and 78.05 ± 4.2%, respectively) and coating onto tanks (58.6 ± 2.1% and 61.2 ± 2.7%, respectively) at 1 mg mL^?1 while least percentage of hatching was recorded in Ag‐TiO₂ nanoparticles irrespective of its concentration & mode of supplementation. Highest survival of L. rohita fry (50.13 ± 2.2%) was observed after 15 days post hatching in CuO coated tanks followed by ZnO coated tanks (38.6 ± 2.8%) while least was recorded in Ag‐TiO₂ coated tanks (22.53 ± 3.0%). However in control tanks coated with Poly‐Urethane base with hardener and uncoated control tanks, the survival was 42.4 ± 1.2% and 41.36 ± 1.8% respectively. Further, significantly lower microbial load of water was recorded in CuO nanoparticles coated tanks (1.5 × 10¹⁰ CFU L^?1) as compared to uncoated control tanks (1.1 × 10¹⁶ CFU L^?1) without affecting water quality parameters. On the other hand, in Ag‐TiO₂ coated tanks, significantly lower microbial load (1.0 × 10⁶ CFU L^?1) as compared to uncoated control tanks at 15 days post hatching was recorded. However, Ag‐TiO₂ was toxic to L. rohita larvae & fry both in direct application and coating onto tanks. Considering the beneficial effects of CuO nanoparticle application, it has the scope of being used in a more eco‐friendly way in hatchery operations.     

Light intensity effects on early life stages of black sea bass, Centropristis striata (Linnaeus 1758)

The effects of four light intensities on growth and survival of first‐feeding stage black sea bass larvae Centropristis striata were investigated in a controlled‐environment laboratory. Fertilized eggs, obtained from LHRHa‐induced spawning of captive broodstock, were stocked (72 eggs L^?1) into twenty 15 L black tanks under light intensities of 100, 500, 1000 and 1500 lx, with five replicate tanks per treatment. The photoperiod was 12L:12D, the temperature was 20°C and the salinity was 35 g L^?1. Larvae were fed rotifers Brachionus rotundiformis from day 2 post‐hatching (d 2ph) at 5–10 rotifers mL^?1. Microalgae Nannochloropis oculata and Isochrysis sp. were added (1:1) daily to maintain a density of 300 000 cells mL^?1. Hatching success and larval growth and survival from d 2ph through d 15ph were monitored. Hatching success was 28–38% under all light intensities, and notochord length at hatching ranged from 2.8 to 3.0 mm, with no significant differences among treatments. By d 15ph, growth (mg wet weight) was significantly higher in the 1000 lx (0.914) and 1500 lx treatments (0.892) than in 100 lx (0.483), and a highly significant trend (P<0.01) towards increased survival with increasing light intensities was observed, from 1.3% at 100 lx to 13.9% at 1500 lx. Higher light intensities within the range of 100–1500 lx improved growth and survival of early larval black sea bass, suggesting that even higher light intensities may improve culture performance. This is consistent with conditions in shallow, near‐shore locations where eggs and larvae are distributed in nature.     

The effect of light intensity on first feeding of the spotted sand bass Paralabrax maculatofasciatus (Steindachner) larvae

The effects of light intensity on feeding incidence and prey consumption at first feeding of spotted sand bass larvae (Paralabrax maculatofasciatus Steindachner), using four light intensity treatments (0, 100, 400, and 700 lx) were evaluated. Specimens were fed the rotifer Brachionus plicatilis at a density of 3 rotifers mL^?1. One hour after the addition of prey, 30±3 (mean±SEM) larvae were sampled from each treatment aquarium. Feeding incidence was evaluated as the percentage of larvae with prey in the digestive tract. Feeding intensity was measured as the number of prey in the digestive tract of the larvae. Histological analysis was carried out to describe the eye structure at the time of first feeding. Larvae fed in darkness (0 lx) had a significantly lower (P<0.05) feeding incidence (1.2±2.2%) and intensity (0.4±0.7 rotifers larvae^?1) than those larvae fed at 100 (28±11%, 1.8±0.2 rotifers larvae^?1), 400 (48±10%, 2.4±0.3 rotifers larvae^?1), and 700 lx (52±4%, 2.4±0.1 rotifers larvae^?1). Feeding incidence of the spotted sand bass larvae increased with light intensity while the feeding intensity showed no significant difference (P>0.05) between light treatments. Histological analysis of the eye structure showed that first feeding larvae had well‐formed lens along with a retina composed of pure single cones as photoreceptors.     

Effect of high dietary intakes of vitamin E and n-3 HUFA on immune responses and resistance to Edwardsiella tarda challenge in Japanese flounder (Paralichthys olivaceus, Temminck and Schlegel)

A feeding experiment was conducted to investigate the effects of high dietary intake of vitamin E (supplied as dl ‐α‐tocopheryl acetate) and n‐3 highly unsaturated fatty acid (n‐3 HUFA) on the non‐specific immune response and disease resistance in Japanese flounder Paralichthys olivaceus. Nine practical diets were formulated to contain one of three levels of vitamin E namely, 0, 80 or 200 mg kg^?1 (the total α‐tocopherol contents in the diets were 21, 97 and 213 mg kg^?1 based on analysis), and at each vitamin E level with one of three n‐3 HUFA levels i.e. 0.5%, 1.5% or 2.0%. Each diet was randomly assigned to triplicate groups of Japanese flounder (initial body weight: 40.5±1.0 g, mean±SD) in a re‐circulation rearing system. Fish were fed twice daily to apparent satiation at 07:00 and 18:00 hours for 12 weeks. During the experimental period, water temperature was maintained at 18±1°C, salinity 31–35 g L^?1, and pH 7.8–8.2. Dissolved oxygen was not less than 6 mg L^?1, and there were negligible levels of free ammonia and nitrite. The results showed that the increase in dietary n‐3 HUFA from 0.5% to 1.0% significantly decreased muscle α‐tocopherol contents in fish‐fed diets with 21 and 97 mg α‐tocopherol kg^?1 diet (P<0.05). In 1.0% HUFA groups, alternative complement pathway activity (ACH₅₀) of fish fed the diet containing the 213 mg α‐tocopherol kg^?1 diet was significantly higher than noted for fish fed the diet containing 97 mg α‐tocopherol kg^?1 diet (P<0.05). Fish fed the diet with 213 mg α‐tocopherol kg^?1 and 2.0% n‐3 HUFA had the highest lysozyme activity (131.7 U mL^?1) among all the dietary treatments. Fish fed the diets containing 97 and 213 mg α‐tocopherol kg^?1 with 1.0% n‐3 HUFA had significantly higher respiratory burst activity than those fed the diets containing 21 mg α‐tocopherol kg^?1 with 0.5 and 1.0% n‐3 HUFA (P<0.05). In the disease resistance experiment, high intake of dietary vitamin E with 213 mg α‐tocopherol kg^?1 significantly decreased cumulative mortality and delayed the days to first mortality after a 7‐day Edwardsiella tarda challenge (P<0.05). In addition, under the experimental conditions, dietary vitamin E and n‐3 HUFA had a synergistic effect on the non‐specific immune responses and disease resistance in Japanese flounder (P<0.05).     

The effects of salinity on reproductive performance and plasma levels of sex steroids in Caspian kutum Rutilus frisii kutum

Sexually mature kutum, Rutilus frisii kutum, captured from its natural habits, the Caspian Sea and the Khoushkrood River, reared at 0.5 g L^?1 and 8–13 g L^?1 for approximately 1 year in experimental condition, for assessing the effect of salinity on reproduction. Plasma concentrations of sex steroid hormones (17β‐estradiol, testosterone and 17α‐hydroxyprogestrone) were measured in the three stages of gametogenesis. Female kutum held at <0.5 g L^?1 or 8–13 g L^?1 had no ovulated oocytes in their ovaries. In contrast, males held in captivity were spermiated, similar to their wild counterparts. The average sperm volume of males held at <0.5 g L^?1 (2.36 ± 0.46 mL) was lower than males held at 8–13 g L^?1 (3.65 ± 0.73 mL) at the end of the experimental period. The highest concentration of testosterone was observed in mid‐gametogenesis in wild fish that was significantly higher than the concentration seen in fish held in either <0.5 g L^?1 or 8–13 g L^?1. Female kutum showed suppressed steroid hormones in captivity, resulting in failure in the gonad development. However, male kutum adapt well to captivity and showed synchrony in steroid hormone variations with the wild fish, resulting in the testicular development. Results of this study also indicate that salinity plays a minor, but vital, role in reproduction of kutum, a factor that needs to be considered for keeping broodstock of brackish water fish species like kutum.     

Methods of microbial control in marine fish larval rearing: clay‐based turbidity and passive larval transfer

This study focused on methods to reduce bacterial loads in the larval culture tanks of California yellowtail (Seriola lalandi). We conducted two trials to evaluate methods to minimize bacterial loads in the larval rearing water. The first trial examined the use of bentonite clay as a turbidity agent to replace algae in a green water‐type environment. This trial consisted of three treatments: (1) clay with continuous feeding (CCO), (2) clay with batch feedings (CBA) and (3) algae paste with batch feedings (ALG). The results showed that both clay treatments had significantly fewer Vibrio colonies in the water column (CBA – 180 ± 78; CCO – 377 ± 120 CFU mL^?1) than the ALG treatment (5692 ± 2396 CFU mL^?1) after 14 days of culture. Survival was significantly higher in the CCO treatment (14.1 ± 2.6%) than either the CBA (2.3 ± 0.5%) or ALG treatments (2.8 ± 1.5%). The second trial attempted to limit bacterial loading in the larval culture tank by passively transferring the larvae into an adjacent, clean tank at 1, 5 and 9 days post hatch during the first 2 weeks of culture. The results from this trial showed that after 12 days of culture, water in the transfertank had fewer Vibrio colonies (1025 ± 541 CFU mL^?1) than the water in the control tanks (1962 ± 1415 CFU mL^?1). Also, survival was significantly higher among larvae that were transferred (43.9 ± 13.5%) than in the control tanks (23.1 ± 6.3%).     

Faecal coliforms in pond water, sediments and hybrid tilapia Oreochromis niloticus×Oreochromis aureus in Saudi Arabia

Total bacterial load, total coliforms faecal coliforms in pond water, sediment, intestine of hybrid tilapia Oreochromis niloticus×Oreochromis aureus and pigeon Columba livia faeces were investigated monthly over a period of 1 year from July 1999 to June 2000. Fish were collected randomly by a cast net. Samples were analysed for coliforms using the multiple‐tube fermentation technique. Results showed total viable bacterial counts in the pond water, sediment, intestine of tilapia and pigeon faeces ranging from 1.8±0.9×10² to 6.0±1.2×10⁴ cfu mL^?1, 3.2±1.2×10⁵ to 2.8±1.5×10⁷ cfu g^?1, 8.2±1.6×10⁵ to 9.9±1.5×10⁷ cfu g^?11.0±0.4×10⁷to9.7±0.2×10⁹ cfu g^?1respectively. The most probable number (MPN) of coliforms and faecal coliforms ranged from 287±12 to ≥1600±0 100 mL^?1 in pond water; the MPN ranges for sediment, tilapia intestine and pigeon faeces were 257±29 to ≥1100±0 g^?1, 237±46 to ≥1100±0 g^?1 and 403±98 to ≥1100±0 g^?1 respectively. The abundance of normal bacteria coliforms was greater in the warm months than in the cold months. Ground water was free from any sort of coliform organisms, and there were no sources of human faecal matter in the pond. So, it is clear that faecal coliforms from pigeon faeces significantly contaminated (P<0.05) the ponds and tilapia intestines. Escherichia coli was the only coliform organism found in pond water, sediment, intestine of tilapia and pigeon faeces.     

Effects of salinity and temperature during incubation on hatching and development of lingcod Ophiodon elongatus Girard, embryos

The interactive effects of salinity and temperature on development and hatching success of lingcod, Ophiodon elongatus Girard, were studied by incubating eggs at four temperatures (6, 9, 12 and 15°C) and five salinities (15, 20, 25, 30 and 35 g L^?1). Hatch did not occur in any of the 15°C treatments. Degree days (°C days) to first hatch was not influenced by temperature or salinity, however, calendar days to first hatch differed significantly for temperature (P<0.0001, 61±1, 44±1 and 35±1 days for 6, 9 and 12°C respectively). Degree days to 50% (427.1±4.2) hatch was not significantly influenced by temperature but was by salinity (P=0.0324). Viable hatch (live with no deformities, 74.1±4.0%) was greatest at 9°C and 25 g L^?1 but not significantly different in the range of 20–30 g L^?1. Larval length (9.4±0.13 mm) was greatest at 9°C and 20–30 g L^?1. Temperature and salinity significantly influenced all categories of deformities with treatments at the upper (12°C and 35 g L^?1) and lower limits (6°C and 15 g L^?1) producing the greatest deformities. The optimal temperature and salinity for incubating Puget Sound lingcod eggs was found to be 9°C and 20–30 g L^?1.