Flavobacteria colonizing the early life stages of hatchery‐incubated Chinook salmon Oncorhynchus tshawytscha (Walbaum 1792) are markedly diverse

Flavobacterial diseases are significant impediments to hatchery‐based fishery conservation and aquaculture productivity worldwide. Recent studies revealed a multitude of novel flavobacteria within the reproductive fluids and unfertilized eggs of feral Chinook salmon Oncorhynchus tshawytscha broodstock, some of which were associated with systemic disease. Herein, embryonated eggs/fry from these broodstock were assayed for flavobacteria while in incubator stacks in three hatcheries over 2 years, as was the water entering hatchery incubators. Overall, >65% of sampled eggs and 38% of fry were colonized by flavobacteria. One hundred and ninety‐one egg and fry‐associated flavobacterial isolates were characterized phenotypically and via 16S rRNA gene sequencing and phylogenetic analyses, revealing that the majority fell into 22 clades (i.e., 15 Flavobacterium spp. groups and seven Chryseobacterium spp. groups) that varied in presence by facility. Although some matched previously described fish‐pathogenic species, the majority were distinct from all described flavobacteria and likely represent novel species. Of concern, iodophor disinfection at the commonly utilized dose/duration for egg‐surface disinfection did not eliminate flavobacteria. Results also implicated maternal routes of infection and source water for some flavobacteria. In total, study findings underscore the complexity of flavobacterial ecology within hatchery environments and highlight the need for improved hatchery biosecurity practices.     

Fry Production of Shire River Tilapia Oreochromis shiranus at Two Broodstock Sex Ratios1

Abstract.— Shire River tilapia Oreochromis shiranus broodfish were stocked at a density of 1.25 fish/m² and sex ratios of either 1:1 or 1:3 (males: females). Two weeks after stocking, fry collection began. Fry were then harvested weekly for 9 wk with a fine-meshed dip net operated from the pond bank for 1.5 h each morning and afternoon. Ten weeks after fry collection began, all ponds were drained and completely harvested. Over the entire 84 d, fry production averaged 0.78 fry/m² per d and did not differ (P < 0.05) between treatments. Fry production per female was significantly higher (P < 0.02) in ponds stocked at a 1:1 sex ratio (111 fry/female) compared to a 1:3 sex ratio (66 fry/female). To avoid inbreeding without sacrificing production, a broodstock sex ratio of 1:1 can safely be used when producing O. shiranus fry at a density of 1.25 fsh/m² in earthen ponds. Competition among females is a possible cause of reduced fry production in densely-stocked brood ponds.     

Water recirculation and good management: potential methods to avoid disease outbreaks with Flavobacterium psychrophilum

Flavobacterium psychrophilum infections cause high mortality among rainbow trout, Oncorhynchus mykiss, fry in Danish fish farms and hatcheries. Hatcheries based entirely on bore‐hole water recirculation systems have been suggested as a possibility for eliminating F. psychrophilum or at least keeping the amount of this bacterium low. The occurrence of the bacterium in a bore‐hole water recirculation system was compared with a combined bore‐hole water and stream water flow‐through system in a hatchery where outbreaks of rainbow trout fry syndrome caused by F. psychrophilum often occurred. Broodfish, unfertilized and fertilized eggs, eyed eggs and fry, as well as water samples from the tanks/troughs with broodfish/fry, were examined. Suspect yellow bacterial colonies were either confirmed or rejected as F. psychrophilum by growth characteristics and by PCR. As both virulent and less virulent F. psychrophilum isolates are known, isolates were characterized. The isolates were ribotyped and grouped according to ribotyping patterns. Representatives of the groups were serotyped. Fry isolates were very homogeneous whereas isolates from broodfish were heterogeneous, whether the isolates originated from external surfaces of the fish (mucus from skin and gills, haemorrhages and ulcers) or internal organs. Flavobacterium psychrophilum was isolated from broodfish in both water systems; 56% of investigated broodfish from the borehole/flowthrough system and 36% from the recirculation facility harboured the bacterium. In the recirculation system, the bacterium was isolated from fish (ulcers, milt, liver, abdominal cavity) kept in the system for 11 months. Flavobacterium psychrophilum was found in milt and ovarian fluid as well as on the surface of fertilized eggs, but not inside the eggs. Fry also harboured F. psychrophilum, but in the water recirculation system the bacterium was first isolated from the fry after they had been graded. Flavobacterium psychrophilum was found regularly in other parts of the hatchery (outside the recirculation facility), including at the time of grading, suggesting that the occurrence of F. psychrophilum in the fry recirculation facility was due to contamination from the borehole/flowthrough hatchery. It is suggested that the combination of bore‐hole water recirculation systems and good management procedures (including egg disinfection) is a possible method for hatcheries to avoid disease outbreaks due to F. psychrophilum.     

Enhancement of the production of turbot, Scophthalmus maximus (L.), larvae by controlling overripening in mature females

Abstract. In turbot, Scophthalmus maximus (L.), daily examination of broodstock females should be a hatchery management practice in terms of fry output. Eggs expelled shortly after ovulation by systematic stripping of females five times a week had high viability (72%) compared to eggs collected twice a week (46%), while handling frequency did not have any effect on female fecundity (total number of ovulated eggs per kg of female). Increase of stripping frequency raised hatching rates and larva production respectively from 11% to 35% and from 20000 to 65000 larvae/kg of spawning female. The enhancement of egg quality and of the resulting larva production are discussed.     

Factors influencing the spawning success of Oreochromis karongae (Trewavas) in ponds

Fry and fingerlings production of a pelagic tilapia from Lake Malawi, Oreochromis karongae subgenus Nyasalapia (Trewavas), was lower (4.07 ± 5.09, 10–21 fry female⁻¹ month⁻¹) than reported for other tilapias (about 135 fry female⁻¹ month⁻¹). Although seasonality was very pronounced in all experimental systems, the use of net bags (hapas) improved fry yield to 24.7 ± 27.90, 0–354 fry female⁻¹ month⁻¹. Multiple regression analysis demonstrated that the depth of ponds, solar radiation, rainfall and maximum temperature significantly influenced fry and fingerling production (adjusted R²= 0.75; P= 0.0024). Injection of exogenous hormones intramuscularly (pimozide, LH-RHa) induced spawning and significantly (P < 0.05) improved spawning success to a maximum of 998 fry female⁻¹ month⁻¹. As in other cultured tilapias, when placed in fine-meshed net bags, broodstock density was critical to fry production. A density of 1.0 fish m⁻² for broodfish of 120–200 g produced significantly larger numbers of progeny in the hatchery system employed. Spawning was seasonal and restricted to November–February. Broodfish ponds given inputs of chicken layers' mash produced significantly (P < 0.05) higher numbers of fry and fingerlings than maize bran (4.17 vs. 1.22 fry m⁻² female⁻¹ month⁻¹).     

Monitoring of the genetic variability of the hatchery and recaptured fish in the stock enhancement program of the rare species barfin flounder <Emphasis Type="Italic">Verasper moseri</Emphasis>

ABSTRACT:   In relation to the stock enhancement program for barfin flounder, hatchery juveniles produced in 2001 were genotyped using microsatellite DNA markers (msDNA) and then released to natural waters. Subsequently, recaptured individuals, designated as 'tentative recaptured', were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, pairwise F _ST and genic differentiation tests were used to estimate the genetic divergence between the wild samples, the hatchery broodstock, and the tentative recaptured samples. Analysis showed significant differentiation among these three groups. Pedigree determination by msDNA was used to establish the origin of the tentative recaptured individuals, in order to elucidate whether they were hatchery produced, 'real recaptured', or wild specimens. Wild individuals were not found. The effective population size of the real recaptured stock was very low ( N _e = 16.6). Equal family survivability was observed between the released and real recaptured stocks, indicating that the genetic variability of the released stock was maintained in the natural environment. Future broodstock management, breeding designs, and family contribution equalization of the offspring to be released will be required in this rare species to avoid unintended genetic differentiation between the wild population and the hatchery broodstock.     

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.     

Improving spawning synchrony in the Nile tilapia, Oreochromis niloticus (L.)

Abstract. The development stage and number of seed (eggs and fry) removed from individual incubating Oreochromis niloticus (L.) broodfish at 10-day interharvest intervals (IHI) was compared with the harvest of free-swimming fry from undisturbed fish held in a tank within the same system. Spawning synchrony and productivity were compared for (1) total exchange of females for ‘conditioned’ females after each harvest, (2) exchange of only spawned females, and (3) return of females back into the spawning tank. Improved synchrony and seed production were positively related to removal of eggs and fry from incubating females and could have important implications for broodstock management in commercial hatcheries. Allowing fish to remain undisturbed in clear water tanks may lead to inhibition of spawning activity related to the formation of hierarchies. Individual weight loss by females during incubation was correlated with stage of seed harvested and thus with timing of spawning during the interharvest interval.     

Genetic assessment of milkfish (Chanos chanos Forsskal) stocks based on novel short tandem repeats for marker‐aided broodstock management

Milkfish hatchery broodstock are either from on‐grown wild‐caught or hatchery‐produced fry/juveniles. To determine if a marker‐assisted management scheme can be formulated for improved milkfish hatchery production, milkfish stocks were genetically characterized using nine novel short tandem repeats or microsatellites. Eight wild‐bred Philippine stocks (CLA, CUR, CAM, SIH, SBH‐I1, HH, PAL and ZH‐P₀), four hatchery‐bred stocks (SBH‐I2, SBH‐D, BoH and ZH‐F₁), two farm stocks of known mixed lineages (SPH and BDH) and one Indonesian hatchery‐bred stock (WJH) were assessed. WJH was included since milkfish fingerlings from Indonesia reared in Philippine farms could be developed into future broodstock. Mean allelic richness (Ar) was highest in wild‐bred stocks (9.5) and lowest in hatchery‐bred spawners (9.1). Mean expected heterozygosities (He) were relatively similar in all stocks with wild‐bred stocks slightly higher (0.67) than the others. An analysis of molecular variance indicated significant yet low genetic differentiation among stocks (F_ST = 0.013; p = .000) where variation (98.6%) was explained by intra‐stock differences. In some of the domesticated stocks, reductions in mean allelic richness were observed in first generation hatchery broodstock (e.g. ZH‐F₁; Ar = 8.3), compared with their founder stock (e.g. ZH‐P₀; Ar = 9.4). The Indonesian stock was similar to local wild‐bred stocks based on genetic variability indices; thus, it might be likely that the local stocks’ fitness traits could be comparable with the imported milkfish stock which has been perceived to be better. The quality of locally available farmed milkfish and prospects of formulating a broodstock management scheme for the production of good quality milkfish seedstock are herewith discussed.     

Early introduction of an inert diet and unenriched Artemia enhances growth and quality of Atlantic cod (Gadus morhua) larvae

The effects of two weaning diets and different weaning protocols on growth, survival, skeletal deformity and gut morphology of Atlantic cod larvae were studied in four groups from 16 to 45 days posthatch (dph). Cod larvae in groups 1 (early weaning with control diet) and 2 (early weaning with experimental diet) were used to evaluate the effects of different polar lipid content of weaning diets on larval and juvenile performance. Cod larvae in groups 2, 3 (early weaning with experimental diet + cofeeding with Artemia) and 4 (earlier weaning with experimental diet and earlier cofeeding with Artemia) were used to evaluate the effects of early introduction of dry diet and Artemia. From 45 to 170 dph, cod juveniles from all four groups were reared using a standard feeding protocol. No significant differences in growth, survival, deformities and gut morphology were found between cod larvae and juveniles from groups 1 and 2. Cod larvae fed on cofeeding regime with Artemia nauplii (groups 3 and 4) were bigger and had lower frequencies of jaw and neck deformities and higher foregut microvillus circumference than cod larvae from group 2. Our results demonstrate the importance of proper weaning protocols in producing better quality cod juveniles.     

Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum), hatcheries: studies on broodstock, eggs, fry and environment

The occurrence of Flavobacterium psychrophilum at four rainbow trout hatcheries was investigated to provide more knowledge about the reservoirs and transmission of this bacterium. Broodstock were sampled at stripping (including both unfertilized and fertilized eggs), and the offspring were then sampled at the eyed egg and fry stages. Water and surface samples (e.g. hatchery trays) were also sampled. Flavobacterium psychrophilum was found in ovarian fluid and milt, indicating that broodstock may serve as a reservoir and are latent carriers of the pathogen. Flavobacterium psychrophilum was not found on or inside eggs, but further egg studies will be necessary to elucidate the possibility of vertical transmission of the pathogen. Flavobacterium psychrophilum was isolated from water samples, but only from water that had been in close contact with farmed rainbow trout or eggs. Flavobacterium psychrophilum isolates were characterized and compared with well-characterized strains, using degradation of elastin, serotype and ribotype profiles. Different ribotypes of F. psychrophilum were found between hatcheries, but a common ribotype A was found at all four hatcheries. Different ribotypes were found in broodstock without clinical disease, whereas only a few ribotypes (mostly ribotype A) were found in diseased fry. The same ribotype A was found in broodstock, in water samples from hatchery trays and in fry, which suggests the possibility of transmission of F. psychrophilum between broodstock and offspring.     

Parentage assignment in hatchery population of brown sole <Emphasis Type="Italic">Pleuronectes herzensteini</Emphasis> by microsatellite DNA markers

ABSTRACT:   Five loci ( Phz2 , Phz6 , Phz7 , Phz12 , and Phz14 ) of microsatellite DNA markers developed in a previous study for parentage assignment in the hatchery population generated by mating among 61 broodstock fish (35 females and 26 males) in a spawning tank, were selected. After natural spawning in the same tank, larvae collected at three different times were categorized into early phase (EP), middle phase (MP), and late phase (LP) groups. In the parental broodstock, the mean number of alleles per locus was 21.8 and expected heterozygosity ( H _E ) was 0.813. In the progeny, the mean number of alleles per locus decreased to 11.6 (EP), 14.4 (MP), and 6.4 (LP) and H _E to 0.796 (EP), 0.833 (MP), and 0.681 (LP). Parental assignment determined eight dams and six sires as major parents for the EP group. In the MP group, 13 dams and ten sires genetically contributed to spawning, but only three dams and two sires were involved in LP group progeny. In the hatchery population produced from a limited number of parental fish such as the LP group, genetic variability was apparently decreased.     

Larviculture of the Mahimahi Coryphaena hippurus in Hawaii, USA

The Mahimahi Coryphaena hippurus has excellent potential for aquaculture due to its fast growth, good food conversion ratio, high fecundity with natural captive spawns, and high price. Using current technology, three crops a year could yield 288,000 kg/ha of water/yr with a potential profit of US $1,280,000/ha water/yr.
Hatchery methods are now adequate for pilot-scale production, and two venture capital companies are pursuing this course. Significant improvement could be made in egg quality, plankton nutrition, plankton substitutes, disease control, reduced aggression, and weaning feeds. More research is needed to learn about digestive physiology, weaning behavior, and the effects of crowding on mahimahi health. Hatchery production is limited most often by the amount of grown out Artemia or yolk-sac mahi larvae provided for postlarval feeding. Problems that occur at the broodstock, hatching, first feeding, second feeding, third feeding, weaning, and early juvenile stages are discussed.
Using the current technology, the cost of post-hatchery mahimahi is 33 cents per fry. This cost could be reduced to 6 cents per fry by successfully (consistently) weaning postlarvae from live feeds by 25 days.     

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.     

To what extent can maternal inherited immunity acquired from a crustacean‐enhanced diet improve the performance and vitality of the offspring and enhance profitability of European Sea bass (Dicentrarchus labrax)?

This study was conducted to investigate the effects of maternal inherited immunity acquired from crustacean‐enhanced diets on the vitality and profitability of sea bass offspring. Newly hatched larvae produced from three groups of broodstock were evaluated. The broodstock were fed (a) a basal diet (BD), (b) a Palaemon‐supplemented diet (PSD), and (c) an Artemia‐supplemented diet (ASD) for 42 days. A total of 400,000 larvae at 3 days posthatch (DPH) produced from each treatment were stocked in larval rearing tanks at 40 larvae/L for 42 days. Survival (%) was improved by 37 and 9.96% in the groups fed ASD and PSD compared with the control group. The growth, swim bladder (%), and condition factor all significantly (p ≤ 0.05) improved in the postlarvae produced from broodstock enhanced with crustacean diets. Compared with the BD group, the serum lysozyme activities of the fish groups fed ASD and PSD increased by 45.6 and 11.7%, respectively. Sea bass fry (90DPH) produced from broodstock fed ASD showed the best tolerance to salinity/temperature stress tests. Furthermore, the profitability improved in ASD and PSD compared with the BD group. In conclusion, sea bass broodstock enhanced with Artemia biomass produced offspring of superior quality with less cost and greater profit margins.     

Evidence of potential vertical transmission of tilapia lake virus

Tilapia lake virus disease (TiLVD) is an emerging viral disease in tilapia with worldwide distribution. Although the horizontal transmission of TiLV has been demonstrated through the cohabitation of infected fish with susceptible fish, no direct experiment showed the potential of vertical transmission from broodstock to progeny. In this study, natural outbreaks of TiLV in broodstock and fry in two tilapia hatcheries were confirmed. The TiLV genomic RNA was detected in liver and reproductive organs of infected broodstock, while infective virus was isolated in susceptible cell line. In situ hybridization assay confirmed the presence of TiLV in the ovary and testis of naturally infected fish and experimentally challenged fish. Moreover, early detection of TiLV in 2‐day‐old fry and the presence of TiLV genomic RNA and viable virus in the testis and ovary suggested the possible transfer of this virus from infected broodstock to progenies. As infective virus was present in gonads and fry in natural outbreak and experimental fish, the importance of biosecurity and prevention of the virus to establish in the hatchery should be emphasized. Hence, the development of TiLV‐free broodstock and the maintenance of high biosecurity standards in the hatcheries are essential for any attempt of virus eradication.     

The Effects of Salinity on Reproductive Performance of Florida Red Tilapia

The reproductive performance of yearling Florida red tilapia broodstock was studied in laboratory aquaria at salinities of 1 (freshwater), 9, 18, 27 and 36 ppt under controlled photoperiod (14 L:10 D) and temperature (28 C).
Spawning was observed at all salinities, although egg production per unit female weight at each spawn was significantly higher at 1 ppt than at higher salinities. An inhibitory effect of salinity on reproductive performance was evidenced by a trend toward lower fertilization, hatching, and survival of prejuveniles with increasing salinity. Fry production per unit female weight declined at salinities above 18 ppt. The results suggest that Florida red tilapia broodstock may be maintained under salinities as high as 18 ppt without impairing fry production, suggesting that hatchery production in brackishwater would be practical in areas where freshwater resources are limiting.     

Genetic structure and genetic relatedness of a hatchery stock of Senegal sole (Solea senegalensis) inferred by microsatellites

Solea senegalensis is a fish species in an early stage of domestication. A commercial hatchery in southern Spain experienced a dramatic failure in spawning and embryo survival, after incorporation into the broodstock of fish reared in their hatchery. To assess the impact of management on the stock quality, a population genetics study was made of adult broodstock present at this hatchery in 2000. The broodstock was composed of a group of fish of wild origin and two more groups from F₁ progeny. A set of eight microsatellite loci was used to compare genetic variability among the three groups and to establish the relationships between pairs of individuals within each group.Individuals from F₁ origin showed a substantial reduction in genetic variability when compared to those from the wild. There was a greater than 50% decrease in the number of alleles per locus corrected by sample size and 16% and 26% reductions in H_e values in the two F₁ groups, respectively. The degree of genetic relationship between individuals, evaluated by the relatedness estimator, rxy, and by graphical approaches, revealed a high proportion, almost 75%, of full-sibs and half-sibs in both F₁ groups, which might account for the reduction in overall variability. The high proportion of siblings also suggests a reproductive pattern in this stock in which few individuals breed successfully. This mating behaviour, along with factors inherent to selection and management systems, might be responsible for the negative effects on the reproductive capacity of the stock.     

Hatchery broodstock conditioning of the blue mussel <Emphasis Type="Italic">Mytilus edulis</Emphasis> (Linnaeus 1758). Part I. Impact of different micro-algae mixtures on broodstock performance

Blue mussel (Mytilus edulis) broodstock collected from the Irish Sea during wintertime (November) was conditioned with three different microalgae diets. Positive flagellates (PF) treatment consisted of Pavlova lutherii, Isochrysis galbana (T-Iso), and Chaetoceros calcitrans (1:1:1). Positive diatoms (PD) treatment consisted of Pavlova lutherii, Chaetoceros calcitrans, and Skeletonema costatum (1:1:1). Broodstock animals in the PF and PD treatments were fed a total of 2.4 × 10¹¹ algae cells per day. Animals in the negative flagellates (NF) treatment received only 1/8th of the total amount of algae of the PF diet. The conditioning diets had an impact on spawning success and broodstock fecundity but not on hatching rate, which was similar in all three groups. The best results were obtained with the PD diet where 84% of the conditioned animals spawned and females released 5.0 × 10⁶ eggs on average. Animals belonging to the PF and NF treatments released, on average, only 3.6 × 10⁶ and 1.6 × 10⁶ eggs, respectively. Although the amounts of algae provided to the broodstock animals had no effect on the hatching rate, the D-larvae resulting from the NF treatment were smaller in size than the larvae from the other treatments. Biochemical analysis of the different broodstock groups at the end of the experiment revealed higher carbohydrate levels in group NF than in PF and PD, supporting the theory that gametogenesis is mainly supported by the energy from the glycogen reserves. As far as we are aware this is the first study describing hatchery broodstock conditioning of blue mussels under fully controlled conditions.