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 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.     

Biochemical parameters of blood plasma and content of microcystins in tissues of common carp (Cyprinus carpio L.) from a hypertrophic pond with cyanobacterial water bloom

The aim of this study was to evaluate the dynamics of the blood plasma parameters and the content of microcystins in the tissues of common carp ( Cyprinus carpio L.) in relation to the toxic cyanobacterial water bloom. Fish (average body mass 2176±697 g) in the hypertrophic pond were exposed to natural water bloom (dominated by Planktothrix agardhii, Pseudanabaena limnetica and Limnothrix redekei ), which contained microcystins (concentration in biomass 20–181 μg g⁻¹ dry wt, concentration in water 0.3–9.5 μg L⁻¹). Biochemical parameters in fish blood plasma were analysed in 89 fish at 14-day intervals during the whole season (nine sampling periods). Our results demonstrated high variability and fluctuations in the investigated parameters. The content of microcystins and density of cyanobacterial cells correlated with some haematological indices as lipase, alanine–aminotransferase, albumin, magnesium and chlorides. The concentrations of microcystins in the muscle and liver of the fish (determined by high-performance liquid chromatography with mass spectrometer) were below the limit of detection during the monitored period [0.31 ng g⁻¹ fresh weight (f.w.) for the liver and 0.13 ng g⁻¹ f.w. for muscle]. Our study demonstrates that although known cyanobacterial toxin microcystins were not detected in the fish tissues, several biochemical parameters important for the fish physiology were modulated by natural cyanobacterial bloom.     

Multiple-dose pharmacokinetic and depletion studies of sarafloxacin in Atlantic salmon, Salmo salar L.

Abstract. Two multiple-dose pharmacokinetic and depletion studies with sarafloxacin hydrochloride in feed pellets were conducted with Atlantic salmon at 12.1 ± 1.1 †C and 9.3 ± 1.5†C, respectively. The dose regimens used were 10mg sarafloxacin kg^-1 biomass for 10 days, and 20mg sarafloxacin kg^-1 biomass for 5 days. In the 10-day study, the highest average concentrations of sarafloxacin in plasma, muscle and liver were 0.14, 0.39 and 0.88μgg(ml)^-1, respectively. In the 5-day study, the highest average concentrations in plasma, muscle, liver and skin were 0.40, 0.61, 1.56 and 0.19μg(ml)^-1, respectively. A comparison between the individual plasma concentrations and the corresponding tissue concentrations revealed significantly higher concentrations in tissue than in plasma during the depletion phase. A similar comparison made in the therapeutic phase from 3 days after first medication to one day after the last medication revealed significantly higher concentrations in liver and muscle than in plasma, and significantly lower concentrations in skin than in plasma. On withdrawal of the drug, sarafloxacin concentrations in plasma and the different tissues declined rapidly, Sarafloxacin was not detected in any plasma sample taken 6 days or more after the end of medication. The corresponding figures for muscle, skin and liver tissues were 14, 20 and 22 days, respectively. The half-lives of sarafloxacin varied in the different tissues included in the studies. The half-life was shortest in plasma, and increased in ascending order in muscle, liver and skin.     

The effect of vitamins C and E in (n-3) highly unsaturated fatty acids-enriched Artemia nauplii on growth, survival, and stress resistance of fresh water walleye Stizostedion vitreum larvae

This study aimed to evaluate the effect of enriching Artemia nauplii with vitamin C (ascorbyl-6 palmitate) or vitamin E (α-tocopherol acetate), 20% w/w, together with a mixture of concentrated eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) on the growth, survival, and stress resistance of fresh water walleye Stizostedion vitreum larvae. Either cod liver oil (CLO) or EPA/DHA ethyl esters concentrate was used as lipid sources in the Artemia enrichment. Walleye larvae were fed ad libitum for 40 days. At day 40, submersion in salt water (25 g L⁻¹) was performed to evaluate larvae resistance to stress. EPA and DHA levels in walleye juveniles fed EPA/DHA-enriched Artemia increased significantly, by an average of 650% compared with fish fed non-enriched Artemia . A significant increase was found for vitamins C (71.8 ± 1.0 and 42.7 ± 1.2 μg g⁻¹ wet weight (WW)) and E (17.0 ± 3.7 and 6.5 ± 0.9 μg g⁻¹WW) concentrations in fish fed enriched and unenriched Artemia , respectively. Growth was comparable throughout treatments, whereas survival was significantly higher in fish fed CLO-enriched Artemia nauplii compared with fish fed Artemia nauplii enriched with EPA/DHA concentrate. The addition of vitamin C increased fish survival by 1.4-fold compared with fish fed Artemia enriched with only EPA/DHA concentrate. The survival of the latter was similar to control fish ( Artemia without enrichment). The supplementation of vitamin E did not affect fish survival significantly. Stress tests revealed that the resistance of walleye larvae to salinity changes increased when Artemia enrichment was supplemented with vitamin C. However, walleye larvae fed CLO-enriched Artemia had the best performances in the stress test.     

Comparative performance of juvenile red abalone, Haliotis rufescens, reared in laboratory with fresh kelp and balanced diets

Juvenile Haliotis rufescens were reared in the laboratory in order to investigate the extent to which fresh kelp and formulated feeds with 250 g kg⁻¹ (25P) and 380 g kg⁻¹ protein content (38P) affected their growth rate, gut residence time (GRT), food consumption ( C ), food conversion ratio (FCR) and digestibility. Abalone from 38P attained the highest growth rate (70.5 ± 4.2 μm day⁻¹; 98.3 ± 6.95 μg day⁻¹), followed by 25P (47.9 ± 2.79 μm day⁻¹; 67.4 ± 2.82 μg day⁻¹) and kelp (23.6 ± 3.36 μm day⁻¹; 28.2 ± 4.11 μg day⁻¹). No significant differences were observed in consumption rate among treatments (0.61–0.68% body weight per day), yet kelp-fed abalone exhibited higher FCR (2.44), protein efficiency ratio (4.42), and apparent digestibility of dry matter (69.5%), protein (69.8%) and gross energy (79.2%) than 38P organisms (59.8, 62.4 and 62.2%, respectively). They also showed longer GRT (23.1 ± 0.93 h). This study demonstrated that formulated diets with 250 g kg⁻¹ and 380 g kg⁻¹ protein inclusion can sustain higher growth rates of juvenile H. rufescens than fresh algae. These differences seem to be due to the amount of dietary protein. Kelp meal appears to improve the consumption and digestibility of balanced diets, and its inclusion in formulated diets is recommended.     

Larval Development and Rearing of Longtooth Grouper Epinephelus bruneus in Jeju Island, Korea

Eggs and sperm were obtained from a female (6.3 kg/BW) and a male (8.4 kg/BW) longtooth grouper Epinephelus bruneus following HCG injection in July 2003. The eggs were fertilized artMcially with the sperm and incubated in one of two 50-m³ tanks after washing the fertilized eggs. The fertilized eggs were 830–950 pn (average 900 ± 2 μm) in diameter and the respective fertilization and hatching rates were 97.7 ± 0.6% and 96.8 ± 0.5% at a water temperature of 25.0 ± 0.5 C. With this regime, the survival rate by day 93 was 7.5% in the 50-m³ tank. The elapsed time from hatching to opening the mouth was 3 d at 25 C. The initial mouth size (z) of the larvae was 0.22–0.23 mm. The newly hatched larvae were 2.02 ± 0.02 mm TL; this increased to 4.12 ± 0.09 mm TL by day 11. By day 54, the larvae had metamorphosed into juveniles and reached 41.12 ± 1.20 mm TL, and by day 93 the juveniles reached 93.78 ± 1.98 mm TL. In all, 49.5% of the larvae were malformed and the type of malformation was diverse.     

Influence of starvation, body size and temperature on ammonia excretion in the marine bivalve Tapes decussatus (L.)

Abstract. Ammonia excretion rate of the marine bivalve Tapes decussatus (L.) varied with body weight, temperature and starvation. There was a steady state in the excretion rates, in which these rates neither increased nor decreased during the first 6 days of starvation. The highest rates of ammonia excreted during the steady state (before decline to lower level) depended on the temperature (715 ± 86 and 395 ± 55 μg NH₄.N/clam/h × 10⁻²) at 28°C and 20°C, respectively. At 16°C, the steady state extended from 6h to 18d starvation. Ammonia excretion rates were higher for starved clams than for fed clams at all sizes, e.g. clams of 0·07 g dry flesh weight (28 ± 9 and 13 ± 5 μg NH₄.N/clam/h × 10⁻² respectively) at 16°C but not at 20°C and 28°C. At each temperature, weight-specific ammonia excretion rates were related to dry flesh weight of starved clams but were not related to fed ones.     

Control of spawning activity in female Nile tilapia (Oreochromis niloticus) (L.) by temperature manipulation

The aim of this study was to maximize the spawning of Oreochromis niloticus females in a specific time period. Females were divided randomly into control and treatment groups. In the treatment groups, females were kept for one week at 28±0.5 °C, after which they were exposed to a reduced water temperature of 22±0.5 °C for 7, 14 and 28 days. Thereafter, the temperature was restored to 28 °C. Females in the control groups were kept continuously at a water temperature of 28 °C. All females were checked daily for signs of spawning for the duration of the experiments and were manually stripped if ready to spawn. The following parameters were calculated for period of 3 and 7 days following a 28 °C temperature restoration: spawning rate, number of eggs per female, weight of female, relative fecundity (eggs g⁻¹ body weight) and the percentage of hatched and swim-up fry. The highest spawning rate of 39.5% was obtained in the 14-day trial over a period of 7 days, while the corresponding value in the control was 12.5%. The percentages of hatched and swim-up fry in the 14- and 28-day trials, however, were significantly higher in the controls than in the corresponding treatment groups.     

Nutritional evaluation of waste date fruit as partial substitute for soybean meal in practical diets of juvenile Nile tilapia, Oreochromis niloticus L.

The potential of waste date meal (WDM; low-quality date palm, Phoenix dactylifera L.) as a carbohydrate source in formulated diets for Nile tilapia was evaluated. Four isocaloric-practical diets (15.7 kJ g⁻¹) were formulated incorporating WDM at 0, 100, 200 and 300 g kg⁻¹ levels as partial substitutes for soybean meal (SBM). These were designated D₀ [284 g crude protein (CP) and 383 g carbohydrate (CHO) kg⁻¹ diet], D₁ (279 g CP and 446 g CHO kg⁻¹ diet), D₂ (207 g CP and 495 g CHO kg⁻¹ diet) and D₃ (175 g CP and 578 g CHO kg⁻¹ diet). Each diet was fed to three replicate groups of 30 fish [20.20 ± 0.09 g (±SE)] for 75 days. No feed-related mortality was observed during the entire experimental period. Final body weight (FBW) and specific growth rate (SGR) in the different treatments were statistically not significantly different ( P  > 0.05). Protein efficiency rate (PER) was lowest in diet D₀ and increased with decrease of SBM content (D₁–D₃). A significant increase in whole body lipid content was recorded in fish fed diets D₂ and D₃. Results showed that WDM could be a substitute for SBM up to 300 g kg⁻¹ in practical Nile tilapia diets without compromising growth.     

Effects of oestradiol-17β or 17α-methyltestosterone administration on gonadal differentiation of largemouth bass Micropterus salmoides (Lacepède)

Monosex populations can be a valuable management tool in culture of larger size largemouth bass (>400 g). In this study, we investigated the effective mode and duration of oestrogen and androgen administrations to produce monosex largemouth bass populations. The experiment consisted of nine treatments. In oral administration groups, we fed 40-day-old fry either 200 mg of an oestradiol-17β (E₂) kg⁻¹ diet or 60 mg of a 17α-methyltestosterone (MT) kg⁻¹ diet for 30, 45 or 60 days. In bath treatments, we immersed fry in a 1 mg MT L⁻¹ solution for 5 h a day on three or six occasions. For control treatment, we fed fry an ethanol-treated diet for 45 days. The frequency of females in the control group was 53.1%. Oral administration of E₂ at all durations resulted in slight increases in the frequency of females (59.8–70.5%). Both modes of androgen administration at all durations were ineffective in altering phenotypic sex. The experimental results of our study indicated that male differentiation passed the point of being completely and functionally influenced by exogenous oestrogens, while female differentiation had already taken place and was no longer responsive to exogenous androgens in 40-day-old (33.5 mm) largemouth bass fry.     

Feasibility of cultivation of Kappaphycus alvarezii (Doty) Doty at different localities on the Northwest coast of India

Kappaphycus alvarezii was cultivated at three places, viz. Mithapur, Okha and Beyt Dwarka on the Northwest coast of India. The biomass and growth rates were measured at 15-day intervals up to 45 days and varied differently for different growth periods at these places. Thus, at 15, 30 and 45 days, the biomass (fresh wt) varied from 182.0±19.96 to 435.0±23.66 g, from 366.0±118.09 to 1096.0±61.43 g and from 530.5±50.95 to 1537.0±43.54 g, respectively, with respective daily growth rates (%) from 3.95±0.78 to 9.80±0.63, from 3.64±0.01 to 13.98±3.07 and from 3.69±0.23 to 6.07±0.06. The seawater temperature, salinity, nitrate and phosphate ranged from 19.3 to 26.9 °C, from 30.81 to 36.83‰, from 11.60 to 19.75 μmol L⁻¹ and from 2.37 to 5.0 μmol L⁻¹ respectively. The growth rate between the culture months was significantly different at Okha at P <0.01. Further, it was significantly correlated to salinity at Mithapur while at Okha and Beyt Dwarka, the same was significantly correlated to nitrate and seawater temperature. Based on this study, commercial cultivation at these localities is quite feasible.     

Effect of different synthetic gonadotrop-releasing hormone analogues and their combinations with an anti-dopaminergic compound on the reproduction performance of sterlet (Acipenser ruthenus L.)

In this study, three synthetic gonadotrop-releasing hormones (GnRH) (azagly-nafarelin; des-Gly¹⁰-( d -Ala⁶)-LH-RH; and des-Gly¹⁰-( d -Phe⁶)-LH-RH) either alone or in combination with metoclopramide were used to induce reproduction of sterlet. The GnRH analogues were applied in a single dose of 40 μg kg⁻¹ of female and 20 μg kg⁻¹ of male body weight. Metoclopramid was administered in a simultaneous injection of 10 and 5 mg kg⁻¹ of body weight for females and males respectively. There were no significant differences in the ovulatory responses of females; ovulation rates varied between 57% and 80%, and at the temperature of 15.5–16.0 °C about 30–34 h were required for final maturation, when eggs of 17.3±1.3% of body weight were stripped. However, the fertilization rates of the des-Gly¹⁰-( d -Phe⁶)-LH-RH-treated groups were significantly lower than that in the other treatment. In males, the combination of the above peptidergic hormones with metoclopramide gave significantly better results than their single application. The results demonstrate that the final stage of gamete maturation in sterlet may be achieved by several hormonal means. The possibility of using new GnRH analogues without dopamine antagonists yields new perspectives for induced breeding of sturgeons, which have particular importance in the light of meat and roe (caviar) production for human consumption.     

Short-term dietary supplementation with the microalga Parietochloris incisa enhances stress resistance in guppies Poecilia reticulata

Two trials were conducted to determine the effects of dietary enrichments with the microalga Parietochloris incisa , rich in arachidonic acid (ARA), on stress resistance in guppies Poecilia reticulata . The microalga was added to commercial diets as a neutral lipid (NL) extract and its fractions or as broken cells. Experimental diets were applied for a period of 14 days. In trial 1, commercial diets were supplemented with NL (containing 25 mg ARA and 0.11 mg β-carotene g⁻¹ feed), its triacylglycerol (TAG) fraction (containing 25 mg ARA g⁻¹ feed and no β-carotene) and the β-carotene fraction (containing 0.11 mg carotenoid g⁻¹ feed and minute amounts of ARA). Neutral lipid-fed fish demonstrated the highest resistance ( P <0.05) to osmotic stress (32-ppt NaCl), followed by fish fed with diets supplemented with TAG and β-carotene alone, which were more resistant than control ( P <0.05). In trial 2, fish fed diets supplemented with higher levels of broken alga (26.1 mg ARA g⁻¹ feed) were more resistant ( P <0.05) to stress as compared with fish fed lower ARA (16.3 mg g g⁻¹) or an unsupplemented control diet. We suggest a dietary supplementation with broken P. incisa cells to enhance stress resistance in guppies before a stressful event.     

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.     

Tissue distribution of vitamin D3 in Atlantic salmon Salmo salar: effect of dietary level

Atlantic salmon Salmo salar were given three dietary doses of vitamin D₃ for a period of 11 weeks: diet groups I, II and III received a 0.04, 2.21 and 28.68 μg g⁻¹ diet of vitamin D₃, respectively. The tissue distribution of vitamin D₃ was investigated, and between diet groups II and III, the level of vitamin D₃ found in the tissues was increased by a factor similar to the increased level in the feed. No effects were observed on weight gain, survival, plasma level of calcium, red blood cell count or haematocrit, relative to the dietary levels of vitamin D₃. The results showed that while the plasma concentration of 25(OH)D₃ increased between diet groups II and III, the concentration of 1,25(OH)₂D₃ decreased.     

Comparison of lipid-walled microcapsules and lipid spray beads for the delivery of water-soluble, low-molecular-weight materials to aquatic animals

Microparticles (< 40 μm diameter) composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were used to encapsulate the low-molecular-weight (mol. wt 460) antibiotic oxytetracycline in the form of either oxytetracycline hydrochloride (OTC.HCl) or oxytetracycline hemicalcium salt (OTC.HEM). Dry, finely ground particles of core material were encapsulated in spray beads. Dissolved core material was encapsulated in lipid-walled microcapsules.
Oxytetracycline (OTC) was most efficiently delivered (≈ 46.5 mg g⁻¹ lipid after 24 h suspension in seawater) as a hemicalcium salt in spray beads. Lipid-walled microcapsules were most efficient for delivering OTC (≈ 8.7 mg g⁻¹ lipid) as OTC.HCl dissolved in 0.2  M HCl at a concentration of 300 mg mL⁻¹.
Spray beads containing OTC.HEM were very stable over 1 month in storage. Lipid-walled microcapsules containing aqueous OTC.HCl lost ≈ 30% of their core material during storage. Freeze-drying of both microparticle types did not improve storage of spray beads, but showed promise for reducing leakage from lipid-walled microcapsules during storage and delivery to suspension feeders.     

Dietary threonine requirement of juvenile hybrid striped bass (Morone chrysops♀×M. saxatilis♂)

Two growth studies were conducted to determine the dietary threonine requirement of reciprocal cross hybrid striped (sunshine) bass. Semipurified diets were prepared with crystalline amino acids and lyophilized fish muscle to supply 350 g crude protein kg⁻¹ diet. The basal diet contained 4.9 g threonine kg⁻¹ from fish muscle, and test diets were supplemented with graded levels of L-threonine. In the first experiment, fish initially averaging ≊ 9.8 g each were fed diets containing threonine levels of 4.9, 7.5, 10.0, 12.5, 15.0 and 17.5 g kg⁻¹ dry diet for 7 weeks. Weight gain, feed efficiency and protein efficiency ratio (PER) were significantly ( P < 0.01) influenced by dietary threonine level. Based on weight-gain responses, a threonine requirement (± SE) of 8.4 (± 0.8) g kg⁻¹ dry diet was determined, and dietary threonine levels of 10.0 g kg⁻¹ diet or greater resulted in the highest levels of free threonine in plasma.
Based on the results of the first experiment, a second feeding trial was conducted with diets containing threonine levels of 4.9, 6.5, 8.0, 9.5, 11.0 and 12.5 g kg⁻¹ dry diet. Fish initially averaging ≊ 3.0 g each were fed each diet for 8 weeks. Weight gain, feed efficiency and PER values of fish were markedly improved, with increases in dietary threonine up to 8.0 g kg⁻¹ dry diet. Regression analysis of weight gain, feed efficiency and PER data using the broken-line model resulted in threonine requirement estimates of 9.7, 8.5 and 8.6 g kg⁻¹ dry diet, respectively. Based on these data, the threonine requirement of juvenile sunshine bass was determined to be ≊ 9.0 g kg⁻¹ dry diet or 26 g kg⁻¹ of dietary protein.     

Induced Spawning of Wild American Shad Alosa sapidissima Using Sustained Administration of Gonadotropin-Releasing Hormone Analog (GnRHa)

American shad Alosa supidissima broodstock were collected from the Susquehanna River during their spawning migration. Mean volume of expressible milt (± standard deviation) was 2.5 (±1.7) mL/kg body weight; mean spermatozoid count was 66.2 ± 10⁹ (±163 ± 10⁹) spermntozoa/mL; and duration of 50% motility was 36.5 (±10.3) see. Ovarian biopsies indicated the presence of oocytes of various sizes (200–2,000 μm in diameter) and stages of development. Fish were implanted with a delivery system loaded with gonadotropin-releasing hormone analog (GnRHa) and started spawning 2 d after treatment. Fertile eggs were collected daily for the next 9 d, for a total of 50,100 eggs/kg body weight with a mean fertilization success of 62%. Upon cessation of spawning, the ovaries of all females still contained large numbers of oocytes at various stages of development, as at the beginning of the experiment, but with a greater number of atretic oacytes. Our observations show that American shad have an asynchronous ovarian development, and treatment with a GnRHa delivery system is effective in inducing several successive spawns of fertile eggs.