Response and condition of larvae of the scallops Nodipecten subnodosus and Argopecten ventricosus reared at the hatchery with different seawater sources

Larvae of the scallops Nodipecten subnodosus and Argopecten ventricosus were reared at a hatchery under five seawater sources: (1) filtered seawater, as the control group; (2) pasteurized seawater; (3) seawater taken from a well; (4) synthetic seawater, and (5) seawater containing a commercial probiotic (Epicin). The quality of each seawater source was measured in terms of counts of Vibrio pathogenic bacteria, levels of nitrites, nitrates, and ammonium, and content of suspended and organic matter. Overall response of larvae under each treatment was measured in terms of growth, survival, biochemical composition, and recruitment rate of spat. Differences in all these parameters, as a function of the seawater source, were analyzed with one-way ANOVA. Larvae survived more, grew faster and larger, had higher protein levels, and recruited more in filtered seawater (in N. subnodosus) and pasteurized seawater (in A. ventricosus), but significant differences between treatments were slight. In A. ventricosus cultures, filtered seawater favored significantly higher Vibrio counts than pasteurized seawater, but this result did not affect the response and condition of larvae. The well seawater treatment ranked third in terms of low bacterial counts, high nitrate levels, larvae showing relatively high survival and growth rates, and spat reaching the settlement stage (only in A. ventricosus). The synthetic seawater and Epicin treatments did not yield satisfactory results for any of the indicators measured. In fact, in the Epicin treatment, larvae survived only the first days of hatchery cultivation and settlement occurred only in A. ventricosus. The results from these first set of trials are useful for planning additional experiments aimed to improve the protocol of seawater use at our research hatchery.     

Comparative study of the culturable microbiota present in two different rearing systems,flow‐through system (FTS) and recirculation system (RAS), in a great scallop hatchery

A comparison of the culturable microbiota present in the different compartments of a great scallop (Pecten maximus) hatchery with two experimental production systems, FTS and RAS, throughout a cycle of larval rearing was carried out. All isolates obtained from broodstock gonads, larvae, tank water, biofilm, microalgae, UV‐treated water and biofilter in both systems at three sampling times were characterized by biochemical tests and identified by sequencing of the 16S rRNA gene. Some of the genera were found to be exclusive of broodstock gonads, such as Sinobacterium, Kordia or Microbulbifer, and differences between gonad microbiota before and after spawning were detected. The number of morphotypes obtained in water, larvae and biofilm was similar in both systems, as well as the behaviour of the microbial populations in almost all hatchery compartments, in which Vibrio, Neptuniibacter, Pseudoalteromonas and Shewanella were the most common genera. The diversity obtained was analysed using the principal component analysis (PCA) and Fisher's exact test, showing that the microbial communities present in the common compartments between FTS and RAS did not significantly differ. These results suggest that, at least from a microbiological point of view, the recirculation system could be a good alternative for the production of scallop larvae.     

Portals of Entry for Gram-Negative Bacteria in a Freshwater Prawn Hatchery

We sought the source of gram-negative bacteria associated with giant freshwater prawns in a hatchery that was previously observed with severe mortality of prawn larvae during the sixth to seventh zoea stages. After thorough disinfection, the gram-negative bacterial microflora of broodstock, larvae, larval rearing tank (LRT) water, and other intake systems were examined for bacteria. Enterobacteriaceae and Vibrio were the major bacteria isolated from the larvae. The present study suggests that Enterobacteriaceae may be introduced via the broodstock and Vibrio via the LRT water.     

Vibrios in hatchery cultures of the razor clam,Solen marginatus (Pulteney)

Hatchery culture of the razor clam, Solen marginatus (Pulteney), has recently been developed in Galicia (NW Spain). However, recurrent episodes of mortalities of larval and post‐larval cultures have been recorded during the course of various studies. The disease signs were similar to those described for other bivalve species in outbreaks caused by bacteria of the genus Vibrio. In this article, we present the results of microbiological monitoring of two batches of razor clams with different survival rates. All fermentative isolates were identified as members of the Splendidus clade within the genus Vibrio. Some of these isolates, identified as Vibrio splendidus‐like, were clearly associated with the batch suffering mortalities, indicating their possible role as pathogens. Similar strains were found in the broodstock, suggesting vertical transmission of these bacteria. This is the first study of the microbiota associated with hatchery culture of S. marginatus, and the results will provide useful information for the optimization of a protocol for hatchery culture of this bivalve species.     

Advances in hatchery and grow-out technology of cobia Rachycentron canadum (Linnaeus)

This paper describes advances in hatchery and grow‐out technology of cobia (Rachycentron canadum, Linnaeus). In 2007, methods for capture, transport, acclimation, sampling, conditioned spawning, larval rearing, fingerling production, nursery, shipping and grow‐out have been perfected. Survival rates ranging from 17.5% to 35% were achieved from egg to shipping size fingerlings (1.0 g) in 2007 at the University of Miami Experimental Fish Hatchery, with production of approximately 20 000 fingerlings per 12 000 L tank. Wild and F1 broodstock cobia have been conditioned to spawn through temperature manipulation producing viable eggs for experimental and production level larval rearing trials in several hatcheries. Brood fish have also been induced to spawn using hormones. Cobia appear to be susceptible to infestations by parasitic protozoa such as Amyloodinium ocellatum and to infections caused by deleterious bacteria such as Photobacterium spp. and Vibrio spp. Prophylactic methods used to prevent and control epizootic diseases at the hatchery are summarized. Improved techniques for cage management were implemented, and both novel designs of submerged cages deployed in exposed areas and traditional gravity cages in protected areas have been used for commercial ongrowing of cobia in the Americas and the Caribbean region.     

The effects of dietary supplements of polyunsaturated fatty acid on pearl oyster,Pinctada margaritifera L., gonad composition and reproductive output

Black‐lip pearl oyster, Pinctada margaritifera broodstock was collected from the wild. Egg production, hatching rate and larval development were compared between oysters induced to spawn within 2 days after collection in the wild (T1), oysters fed a pure microalgae diet during 24 days before spawning (T2) and oysters fed the same microalgal diet in which 10% of the algae were replaced with 2 μm polyunsaturated fatty acid (PUFA)‐rich microspheres (T3). Administration of lipid microspheres resulted in larger sized eggs, a higher percentage of D‐larvae and larger sized 48‐h‐old larvae (P<0.05). The total and neutral lipid contents of the gonad increased after oysters were fed with microalgae only or with supplementary diet. The major neutral and polar fractions of saturated fatty acid (SFA) were 16C and 18C fatty acids, and not influenced by the diet (P>0.05). The gonads of oysters fed supplementary PUFA contained more docosahexaenoic acid (DHA) and less monounsaturated fatty acids. Higher level of DHA in gonads of T3 was associated with oogenesis and embryogenesis success. The n‐3/n‐6 ratio in the neutral lipid fraction provides a good indication of the spawning condition and predicting egg size and hatching rate.     

Preliminary study on broodstock rearing,induced breeding and grow‐out culture of the sea cucumber Holothuria scabra in Sri Lanka

Experiments were conducted to identify suitable methods for broodstock rearing, induced breeding and grow‐out culture of Holothuria scabra in Sri Lanka. Two hundred and seventy‐two brooders (500–600 g) collected from off Mannar were individually packed in oxygen‐filled polythene bags with and without sea water and transported to a sea cucumber hatchery at Kalpitiya. Lagoon pens, sand‐filled fibreglass tanks and bare tanks were used in triplicates to maintain brooders. Spawning was initiated using air dry, water jet and thermal‐stimulation methods. Hatchery produced juveniles with an average weight of 11 ± 5 g were reared (2 individuals m^?2) in lagoon pens, mud ponds and fibreglass tanks in triplicates. The significantly high evisceration rate was observed when brooders were transported without sea water (t‐test, P < 0.05). Brooders maintained in bare tanks showed a significant weight reduction than the brooders in sand‐filled tanks and lagoon pens (anova , P < 0.05, d.f. = 2). Thermal stimulation (ambient temperature ± 3–5°C) was found to be the most successful method of spawning initiation of H. scabra. The mean (±SD) percentage males and females participated for spawning per trial was 9.2 (±10) and 4.6 (±5.6) respectively. On an average, 1.16 millions of eggs (±1.03 SD, n = 5) were obtained per spawning trial. H. scabra juveniles reared in tanks showed significantly lower growth rate than the juveniles in pens and ponds (anova , P < 0.05). Lagoon pens and sand‐filled tanks are suitable to maintain brooders and lagoon pens can be successfully used for mass rearing of juveniles.     

Proximate and fatty acid composition of the gonads of wild versus hatchery‐conditioned Pinctada margaritifera broodstock

The composition of protein, carbohydrate, lipid and fatty acids of the gonad of wild female broodstock of black‐lip pearl oyster, Pinctada margaritifera, was compared with oysters fed on a ternary combination of microalgae in hatchery. Artificial feeding was found to be as good as natural feeding in terms of number and size of released eggs. Lipid, protein and carbohydrate reserves of unfed oysters were found to be insufficient to complete oogenesis. The proportions of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) of the neutral and polar lipids extracted from female gonads were not influenced by variations in the fatty acid composition of the natural food and ternary combination of microalgae in hatchery. T‐Iso, Chaetoceros calcitrans and Chaetoceros muelleri were able to provide sufficient 22:6n‐3 (DHA) and 20:5n‐3 (EPA), two of the most important essential fatty acids required for gametogenesis. The n‐3/n‐6 and 22:5n‐3/20:4n‐3 ratios were consistently higher in the neutral lipids than in the polar lipids. Conversely, the ratio of 20:4n‐3/20:5n‐3, 22:6n‐3/20:5n‐3 and PUFA/SFA was higher in the polar lipids.     

Enhancement of <Emphasis Type="Italic">Penaeus monodon</Emphasis> shrimp postlarvae growth and survival without water exchange using marine <Emphasis Type="Italic">Bacillus pumilus</Emphasis> and periphytic microalgae

We have investigated the possibility of using a consortium of marine bacterium and periphytic microalgae to improve the water quality and increase the growth and survival of the shrimp Penaeus monodon in a hatchery system. Three treatments were evaluated for their effect on P. monodon postlarvae (PL) when the culture water was not changed: Bacillus pumilus alone (B); periphytic microalgae alone (M); B. pumilus + periphytic microalgae (BM). P. monodon PL raised in a tank of unchanged water without bacterium and periphytic microalgae served as the control. The water in tanks of the M and BM treatments had significantly low levels of total ammonia-nitrogen (TAN) (0.03 and 0.01 mg l⁻¹, respectively) and nitrite-nitrogen (NO₂-N) (0.03, 0.01 mg l⁻¹, respectively) than that in the B (TAN 0.80, NO₂-N 0.68 mg l⁻¹) and control (TAN 1.11, NO₂-N 1.12 mg l⁻¹) tanks. Moreover, PL cultured in tanks M and BM had significantly higher survival and specific growth rates and a significantly higher resistance to the reverse salinity stress test than those in the B and control tanks. Compared to the control PL, the PL cultured in the BM tanks had significantly higher levels of protein, lipid, polyunsaturated fatty acids, ecosapentaenoic acid, and docosahexaenoic acid. The culture water in tanks BM also contained significantly less Vibrio than the control water. Our results illustrate the beneficial effects of a B. pumilus and periphytic microalgae consortium on improving the water quality and the growth and survival of shrimp PL grown in a hatchery system.     

Quantitative and Qualitative Aspects of Bacterial Communities Associated with Cultures of Chlorella minutissima

Microbiological conditions in 200 L cultures of microalgae Chlorella minutissima in polyethylene sleeves were examined. The influence of addition of antibiotic (nitrofurantoin) was studied. Samples were taken 2, 5, 9, and 14 d after inoculation and were spread on solid Zobell medium and on thiosulfate citrate bile salts sucrose agar for the enumeration of total numbers of bacteria and numbers of presumptive Vibrio bacteria, respectively. Microalgae grew well in the cultures and high numbers of culturable bacteria (about 10⁷ bacteria/mL) were present in all microalgae cultures. Addition of antibiotic in 200‐L cultures of C. minutissima did not result in a significant decrease of total counts of bacteria per unit volume (P > 0.05), while the numbers of presumptive Vibrio were higher in cultures added antibiotic compared with the control treatment (P < 0.05). The numbers of presumptive Vibrio per unit volume and the percentage of fast‐growing bacteria were higher in microalgae supernatant than in noncentrifuged samples. Representative colonies were taken from all samples and in total 649 bacterial strains were isolated during this study. A range of phylotypes was identified by amplification and sequencing of the 16S rDNA gene and phylogenetic trees were constructed. Several members of the Roseobacter clade were dominant among the culturable isolates.     

Effect of different broodstock densities on the reproductive performance of Nile tilapia, Oreochromis niloticus (L.), in a recycling system

The present study was conducted to evaluate the effect of different stocking densities on the seed production of Nile tilapia, Oreochromis niloticus (L.), under intensive recycling hatchery system conditions. Males and females with mean body weights of 163.2 and 105.0 g, respectively, were stocked at three broodstock densities (4, 8 and 12 fish m^–2) at a male:female ratio of 1:3 in 1 × 1 × 0.43 m (W × L × H) fibreglass tanks. The tanks were illuminated at 2500 lux for 18 h day^–1 and the water temperature was maintained at 29 ± 1 °C. Effluent from spawning tanks was recycled through a biological filter with 10–15% replacement of new water per day. The experiment lasted for 126 days. The results showed that breeders stocked at 4 fish m^–2 had significantly higher (P < 0.05) mean values for total seed production, seed kg^–1 female day^–1, seed female^–1 day^–1, seed m^–2 day^–1 and spawning synchrony than at 8 and 12 fish m^–2 broodstock densities. The mean percentage of seeds in the yolk-sac and swim-up fry stages was highest at 4 fish m^–2 broodstock density. However, the recovery rate was not affected by broodstock density. It is recommended that further research should be conducted to determine whether weight m^–2 , number m^–2 or age of broodstock should be the basis for stocking broodstock.     

Effects of different diets and seawater systems on egg production and quality of the broodstock Babylonia areolata L. under hatchery conditions

The broodstock of spotted Babylon snail, Babylonia areolata, were conditioned in indoor tanks under two seawater systems (static system and flow‐through system) and five types of foods (fish, shrimp, squid, green mussel and formulated diet). High survival and good feeding was found from broodstock conditioned in all treatments of seawater systems and diets. All broodstock showed active behaviour with no signs of stress as exhibited by active movement and feeding, protrusion of siphonal tube and constant egg laying throughout the experiment. Two‐way analysis of variance showed that there were no significant differences (P>0.05) in the total yield of egg capsules, size of egg capsules, size of newly hatched larvae and growth of larvae among broodstock B. areolata conditioned in the seawater system and diet treatments, but there were significant differences (P>0.05) in the frequency of egg capsule laying, number of fertilized eggs/embryos per capsule, total yield of fertilized eggs/embryos and final survival of larvae among broodstock B. areolata conditioned in the diet treatments. We present initial evidence that the seawater system had a stronger influence on the egg production and quality of broodstock B. areolata than conditioning diets.     

Observations on growth, sexual maturity and spawning performance of pond-reared Penaeus merguiensis

This 12‐month preliminary study investigated the development of sexual characters, primary sexual maturity, ovarian maturity and spawning performance of pond‐reared Penaeus merguiensis in relation to culture conditions in south‐east Queensland, Australia. Post‐larvae of P. merguiensis were produced and cultured in two 60‐m³ tanks during the first 14 weeks. Before winter, they were harvested and stocked in three different overwintering facilities: a 200‐m² covered pond, two 60‐m³ outdoor tanks and a 15‐m³ indoor, recirculated tank at a stocking density of 10 individuals m^?2. The development of sexual characters was found to be similar to that reported previously in wild P. merguiensis. Males matured at younger ages and smaller sizes than females. Overall, the average size at primary sexual maturity of pond‐reared P. merguiensis was 23.1 mm carapace length for males (possessed spermatophores) and 29.3 mm for females (being impregnated). Water temperature and the availability of natural food strongly influenced prawn growth, maturity rate and their subsequent spawning performance. Growth, maturity rate and spawning performance of prawns in the covered pond were significantly higher than in the other overwintering facilities. Prawns started mating at 6–7 months, reached full ovarian maturation and spawned as early as about 8 (peaked at 9–11) months from hatching, producing high fecundity and viable larvae. There was a strong relationship (P < 0.001) between prawn size and fecundity. The results of this study suggest a potential for using pond‐reared broodstock P. merguiensis for hatchery production and for domestication or selective breeding programmes.     

The effect of temperature and conditioning interval on the spawning success of wild-caught blacklip (Haliotis rubra, Leach 1814) and greenlip (H. laevigata, Donovan 1808) abalone

Wild‐caught blacklip (Haliotis rubra, Leach 1814) and greenlip (H. laevigata, Donovan 1808) abalone fed a formulated feed were held at 16 or 18°C for different conditioning intervals ranging from 114 to 235 days and induced to spawn using ultraviolet‐irradiated seawater. They were conditioned again for a second identical period before another induction. For H. rubra, mean spawning rate of both sexes was higher in groups held at 18°C than at 16°C, as was the repeat spawning rate. Conversely, animals held at 16°C produced significantly more gametes than those at 18°C. Egg production peaked in groups held at 16°C for ≥165 days. While both mean and total sperm production of H. rubra varied significantly, both figures were always high. Unlike H. rubra, the spawning rate, repeat spawning rate and gamete production of both sexes of H. laevigata were higher when cultured at 16°C than at 18°C. Egg production peaked in groups conditioned at 16°C for ≥212 days. Both mean and total sperm production by H. laevigata were much lower than for H. rubra. This study demonstrates that year‐round hatchery production of seedstock of both species is possible providing broodstock are held under favourable environmental conditions, preferably 16°C.     

Enhancement of reproductive performance in shrimp Litopenaeus vannamei (Boone, 1931) by supplementation of Ulva clathrata meal in maturation diet in two commercial hatcheries

The effect of supplementing dehydrated seaweed Ulva clathrata (20 g/kg) in the squid fraction of a formulated fresh diet on the reproductive performance of Litopenaeus vannamei broodstock was evaluated in two commercial hatcheries (A and B). The origin of broodstock, use of ablation on females, tank/culture conditions (for maturation, spawning, and larval rearing) were different between hatcheries. Daily mortality was higher (p < .01) for females in the control group compared to Ulva‐fed group of hatchery A, where females were ablated. No differences in mortality were found for unablated females in hatchery B, or with males from either hatchery. In hatchery A, broodstock fed Ulva produced more eggs (an increase in +5% compared to control, p < .05) and nauplii per female (+18%, p < .01). In hatchery B, broodstock fed Ulva produced significantly more eggs (+26%, p < .05) and nauplii per day (+33%, p < .05). In hatcheries A and B, eggs from broodstock fed Ulva had increased hatching rates (+13 and +7%, respectively, p < .001 in both cases). The number of spawns per day had a tendency to be higher in broodstock fed Ulva in both hatcheries, but the difference did not reach statistical significance (+14% for hatchery A and +20% for hatchery B). Dehydrated U. clathrata appears therefore as a functional additive in L. vannamei maturation diets that enhances the reproductive performance at commercial scale, regardless of the management conditions.     

Sustained, Natural Spawning of Southern Flounder Paralichthys lethostigma Under an Extended Photothermal Regime

Hormone‐induced spawning of southern flounder Paralichthys lethostigma has produced substantial numbers of viable eggs, but wide variations in fertilization and hatch rates have been reported. Recently, sustained natural spawning of southern flounder broodstock, without hormone induction, has been achieved in our laboratory. Adults (average weight = 1.12 kg; N= 25), including 6 captured as juveniles in 1993 and 19 captured as adults during September 1998, were stocked in two 4.8‐m³ controlled‐environment tanks in October 1998 and held under natural photothermal conditions until January 1999, when an artificial winter photo‐period of 10 L:14 D was initiated and then maintained through April 1999. Sex ratio was approximately 13 females:8 males:7 unknown. Natural spawning was observed in early December 1998 and increased in frequency to a peak in March 1999, before declining in late April. Water temperature ranged from 13.9 to 24.5 C during the spawning period. Natural spawnings over 142 d produced a total of 18.3 × 10⁶ eggs, with a mean fertilization rate of 28.0% (range = 0–100%), yielding 4.94 × 10⁶ fertilized eggs. The mean percentage of eggs that remained buoyant in full‐strength seawater (34 ppt) was 41.3% (0–98%), while hatching rate of buoyant eggs was 37.3% (0–99%) and survival of yolksac larvae to the first‐feeding stage was 30.2% (0–100%). Gonadal biopsies in late April identified six females from both tanks as probable spawners. A preliminary comparison suggests that natural spawning produced much larger numbers of viable eggs per female, with higher egg quality (i.e., fertilization and hatching success) than hormone‐induced spawning. In contrast to natural spawning, hormone‐induced strip‐spawning enabled timing of spawnings to be more precisely controlled. These results suggest that a combination of both natural and hormone‐induced spawning of photothermally conditioned fish will help produce the large numbers of eggs required to support commercial production.     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.     

Notes on hatchery spawning methods for bigmouth sleeper Gobiomorus dormitor

The bigmouth sleeper Gobiomorus dormitor, a popular sport fish within its native range, offers a management alternative to exotic species currently managed for recreational fishing. Hatchery production and stocking are needed to create reservoir sport fisheries because this species usually requires access to marine environments for recruitment. Bigmouth sleepers have not been spawned previously in a hatchery, and hence we used natural, artificial and semi‐natural spawning techniques. No egg deposition or propagation resulted from natural pond spawning. Artificial spawning techniques using Ovaprim^® and Chorulon^® injections followed by hand stripping were more successful. Semi‐natural (hormone injections plus volitional spawning) spawning trials using Ovaprim^® and Chorulon^® displayed the greatest potential, and three consecutive evening injections (1.0 mL kg^?1) of Chorulon^® appeared most conducive. High‐fecundity spawning was achieved by injecting bigmouth sleeper pairs and allowing volitional spawning in aquaria with spawning cavities. Initial success in 2007–2008 was isolated to wild‐caught fish, but captive broodstock were spawned using these techniques in 2009. Eggs hatched in <20 h at 23–24 °C. The resulting larvae, 1.0–1.5 mm in length, displayed large yolk sacs, unpigmented eyes and no apparent mouth. These hatchery propagation efforts suggest that hormone‐induced semi‐natural spawning has the greatest potential for bigmouth sleeper propagation.     

Spawning of the Temperate Sea Cucumber,Australostichopus mollis (Levin)

The spawning of sea cucumbers is related to the interaction of environmental cues and reproductive maturity. Groups of Australostichopus mollis were collected every 2 wk during the summer reproductive season from mid‐October to early February. They were observed for spawning in the hatchery during heat‐shock trials conducted 3–5 C above ambient seawater temperature. Lunar periodicity existed with spawning being most prominent in the week following a full moon. In the week prior to a spawning event, there was also a decrease in ambient incoming seawater temperature. More than 75% individuals spawned during most trials in which spawning occurred. In total, 40% of the 270 individuals spawned during the reproductive season. Reliable spawners were females with a yellow gonad color and large numbers of vitellogenic oocytes, and males with large numbers of spermatocytes and an absence of motile spermatozoa. Furthermore, the percentage germinal vesicle breakdown of oocytes exposed to starfish radial nerve extract compared to seawater was greater for the more orange‐colored gonad of nonspawners. Gonad color is useful in determining the likelihood of spawning over a 2‐wk period in the presence of a cue. Sea cucumbers held for short periods in the hatchery during the reproductive season maintain a natural rhythm of spawning that can be monitored using visual indices and augmented using heat shock.     

Vertical transmission and concurrent infection of multiple bacterial pathogens in naturally infected red tilapia (Oreochromis spp.)

Horizontal transmission of various bacterial pathogens in tilapia is well described, but there is scant information regarding their vertical transmission. This study aimed to determine the possibility of vertical transmission of two commonly reported bacterial pathogens (Francisella noatunensis subsp. orientalis and Shewanella putrefaciens) in natural stocks of red tilapia (Oreochromis spp.). Vertical transmission of these pathogens via gametes was evaluated using in vitro fertilization from 10 different families and analysing for the presence of bacteria in milt, unfertilized eggs, fertilized eggs and offspring at various ages (1‐day‐old larvae, 10‐day‐old fry and 30‐day‐old fingerlings), as well as water samples using colorimetric LAMP assay. The study revealed that both F. n. orientalis (6/10) and S. putrefaciens (4/10) was transmitted vertically to the fertilized eggs. Analysis of the water samples from different water sources (brood stock tanks, hatching chamber and larval rearing tanks) showed that both the pathogens were present in water samples with highest prevalence for F. n. orientalis followed by S. putrefaciens. Analyses for the presence of two pathogens in various organs (gonads, gill, liver, spleen, kidney and brain) of the healthy tilapia broodstock without any clinical symptoms of disease demonstrated they were carriers of S. putrefaciens and F. n. orientalis. This is the first documented evidence that vertical transmission via the broodstock of tilapia may also play an important role in transmitting these problematic pathogens to their progeny and underlines the necessity to modify the current disease management strategies in tilapia aquaculture.