Streptococcus iniae infection and tissue distribution in hybrid striped bass (Morone chrysops×Morone saxatilis) following inoculation of the gills

This study was designed to test the possibility that Streptococcus iniae enters through the gills and causes infection in hybrid striped bass. To determine the dose response, four groups of fish were inoculated with S. iniae via the gills with a dose of 5.0×10⁵, 2.6×10⁶, 5.0×10⁶, or 1.0×10⁸ CFU/fish. One group of fish was inoculated with tryptic soy broth (TSB) via the gills to serve as controls. The cumulative percent mortality was 13%, 27%, 100% and 100% for 5.0×10⁵, 2.6×10⁶, 5.0×10⁶ and 1.0×10⁸ CFU/fish, respectively. We also examined the tissue dissemination of S. iniae at 0.5, 4, 8, 12, 24, 48 and 72 h after experimental gill inoculation. Fish were inoculated with 2.6×10⁶ or 5.0×10⁶ CFU/fish, which caused low and high mortality, respectively. Within 48 h, fish inoculated with the 2.6×10⁶ dose were culture positive on the gill surface, blood of the first and second gill arches, blood of the third and fourth gill arches and the nares. However, for the dose of 5.0×10⁶ CFU/fish, S. iniae was also isolated from the olfactory, optic and cerebellum regions of the brain, eye, head kidney, trunk kidney, spleen and liver at 48 h. For the 2.6×10⁶ dose, S. iniae was not isolated until 48 h post-inoculation, but was isolated at 12 h for the 5.0×10⁶ dose. The results of this study indicate that S. iniae can enter hybrid striped bass through the gills. However, mortality at similar S. iniae doses was lower than we previously observed by inoculation of the nares.     

Safety and efficacy of emamectin benzoate administered in-feed to Atlantic salmon, Salmo salar L., smolts in freshwater, as a preventative treatment against infestations of sea lice, Lepeophtheirus salmonis (Krøyer)

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg⁻¹ fish day⁻¹ for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg⁻¹ day⁻¹, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg⁻¹ day⁻¹. Among smolts that received 272 μg kg⁻¹ day⁻¹, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified.

In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg⁻¹ day⁻¹ (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P<0.001) on treated fish than on controls. When fish challenged at day 109 were sampled at day 128, efficacy was 44.3%, but survival of chalimus to adult lice on treated fish was lower, and at day 159, efficacy had increased to 73%. These results demonstrate that treatment of salmon smolts with emamectin benzoate in freshwater was well tolerated and highly effective in preventing sea lice infestation following transfer of fish to seawater.     

Effect of water temperature on the immune response of white shrimp Litopenaeus vannamei to Vibrio alginolyticus

White shrimp Litopenaeus vannamei held in 25‰ seawater at 27 °C or 28 °C were injected with TSB-grown Vibrio alginolyticus at 1 × 10⁴ colony-forming units (cfu) shrimp^− 1 or 1 × 10⁵ cfu shrimp^− 1, and then cultivated onward at water temperatures varying from 20 to 34 °C. Over 24–144 h, mortality of V. alginolyticus-injected shrimp held at 34 °C or 32 °C was significantly higher than that of shrimp held at lower temperatures. In a separate experiment, shrimp held in 25‰ seawater at 28 °C and then cultured onward at 20 to 32 °C were examined for immune parameters at 24–96 h. THC, phenoloxidase activity, respiratory burst, and SOD activity decreased significantly at 24 h after transfer to 32 °C. Shrimp held in 25‰ seawater at 27 °C and then cultured onward at 20 to 34 °C showed a significant reduction in phagocytic activity and clearance efficiency for V. alginolyticus at 24 h after transfer to 34 °C. It was concluded that transfer of shrimp from 27 or 28 °C to higher temperatures (32 and 34 °C) reduced their immune capability and decreased resistance to V. alginolyticus infection.     

Bacterial diversity of tilapia (Oreochromis niloticus) cultured in brackish water in Saudi Arabia

The bacterial flora occurring in brackish pond water, sediment, gills and intestine of healthy tilapia cultured in Saudi Arabia were estimated both quantitatively and qualitatively, and the isolates were identified to genus or species level. Total viable count of bacteria ranged from 1.4±1.5×10³ to 8.6±2.7×10³ cfu ml⁻¹; 1.2±3.1×10⁶ to 7.3±1.1×10⁷ cfu g⁻¹; 8.7±1.9×10⁵ to 2.1±0.9×10⁶ cfu g⁻¹; and 2.8±2.4×10⁷ to 1.0±1.6×10⁸ cfu g⁻¹ in the pond water, sediment, gills and intestine of brackish water tilapia, respectively. In total, 19 bacterial species were identified. The bacteria were predominantly Gram-negative rods (87%). Pond water and sediment bacteria influenced the bacterial composition of gills and intestine of tilapia. In contrast to gill bacteria, more diversification was observed in intestinal bacteria. The predominant (prevalence >10%) bacterial species were Vibrio parahaemolyticus, Vibrio carchariae, Vibrio alginolyticus, Chryseomonas sp., Vibrio vulnificus, and Streptococcus sp. in all the populations with the exception of the sediment population where Streptococcus sp. was replaced by Shewanella putrefaciens. Vibrio spp. (58% of the total isolates) dominated the total bacterial population.     

Infection and mortality by the yeast Metschnikowia bicuspidata var. bicuspidata in chinook salmon fed live adult brine shrimp (Artemia franciscana)

First-feeding chinook salmon were fed either live adult Artemia franciscana or commercial feed over a 15-week period. Unexplained mortality began occurring in the Artemia-fed fish after 35 days on the diets, with cumulative mortality reaching 34.5% in the Artemia-fed fish compared with 4.3% in feed-fed fish. Necropsy examinations revealed systemic fungal yeast infections and the causative agent was cultured from kidneys. Fungal cells were observed in the transport water of Artemia shipments and within the Artemia. The Artemia were purchased from a retail supplier, and originated from salt ponds in San Francisco Bay (SFB), California. Artemia infection rates ranged from 0.5% to 37.5% (mean 16.5±3.59%) in shipments received over a 5-week period. The fungus was characterized by morphological and physiological properties and was identified as Metschnikowia sp., a pathogenic yeast of aquatic invertebrates. The yeast grew at 9–27 °C, and 0–180 ppt NaCl, and could tolerate salinity of at least 270 ppt. Sequence analysis of the divergent D1/D2 domain of ascomycetous yeast 26S ribosomal DNA identified the organism as Metschnikowia bicuspidata var. bicuspidata. The organism was found in water from the salt ponds and probably entered from the bay. Mortality due to the fungal infection stopped after changes were made in the handling of incoming Artemia shipments.     

Life cycle of Caligus rogercresseyi, (Copepoda: Caligidae) parasite of Chilean reared salmonids

Caligus rogercresseyi, [Contrib. Zool. 69 (2000) 137] is the only caligid known to affect the salmon industry in Southern Chile. Economic losses due to reduced fish quality, cost of chemical treatment and outbreaks of other diseases such as the Piscirickettsiosis occur. The life cycle of C. rogercresseyi is described in rainbow trout reared in seawater tanks from observations made under natural conditions of light and temperature between January 1997 and April 1998. Fish were infected with laboratory-cultured larvae obtained from ovigerous females. Rainbow trout were periodically slaughtered for parasite collection and identification. C. rogercresseyi life cycle includes the following stages: two nauplius, one copepodid, four chalimus and the adult. No preadult stage was observed. Timing of the different stages of development was directly dependent on water temperature. The maturation of the eggs or the time for a complete life cycle took place at 45 days in July at 10.3 °C, 31–32 days in April at 12.4 and 12.8 °C, respectively, and at 26 days in November at 15.2 °C. In January, at 16.7 °C, only the appearance of first eggs were observed at 18 days. A simple degree–day (dd) model is proposed for each developmental stage between 4 and 17 °C, where the development rate is a linear function of the average temperature of water. Using this degree–day model, the proportion of fourth stage chalimus was maximum at 172 dd of effective temperature, adult males at 193 degree–days, adult females at 208 dd. The minimum temperature threshold is at 4.2 °C where there is no development of the parasite. The appearance of first eggs occurred at 231 dd and the first pigmented eggs at 277 dd. The temperature-independent degree–days value allowed to predict the timing of C. rogercresseyi life cycle at any temperature within the evaluated range.     

In vivo fish models for visualizing Aeromonas hydrophila invasion pathway using GFP as a biomarker

The invasion pathway of Aeromonas hydrophila in vivo was studied in Crucian carp (Carassius auratus gibelio) with a virulent strain A. hydrophila J-1 transformed with a plasmid encoding green fluorescent protein (pGFPuv) (A. h J-1^GFP), which had similar virulence characteristics (haemolysin, extracellular proteases production, toxicity for EPC cells, survival in fish serum and LD₅₀ value) as the parent strain. Fish were divided into four experimental groups: (1) normal fish; (2) bacteria bath challenged, unwounded fish; (3) skin artificially wounded by scalpel, bacterial bath challenged fish; and (4) skin mucus layer partially removed by paper towel, bacterial bath challenged fish. The number of bacteria from blood, gills, kidney, muscle, liver and intestine were detected at 2, 4, 8, 12, and 24 h post-challenge. High bacterial numbers were observed in the muscle of the artificial wound group, in the kidney of the mucus removed group and in the gills of all groups. In conclusion, the gills and damaged skin are likely to be the main routes of entry for A. hydrophila, and GFP can be used as a real-time biomarker to study intimate host–pathogen interaction in fish.     

Beneficial effects of n-3HUFA enriched Artemia as food for larval palmetto bass (Morone saxatilis × M. chrysops)

The beneficial effects of feeding n−3 highly unsaturated fatty acids (HUFA ≥ 20 carbon fatty acids with three or more double bonds) to palmetto bass (striped bass × white bass) larvae, 4–30 days of age, were studied using Artemia diets enriched with six n-3HUFA levels. Dietary n-3HUFA concentrations were < 0.03% (control diet), 0.33%, 0.63%, 0.87%, 1.26%, and 2.27% of dry-wt Artemia. Larval n-3HUFA contents were reduced at a faster rate with decreasing dietary n-3HUFA intake, and were significantly different by 30 days posthatch (4–20 mg g⁻¹ dry-wt fish). Starved larvae selectively conserved endogenous n-3HUFA reserves, indicating an essential role of n-3HUFA in larval development. Mass mortality in the control and 0.33% n-3HUFA diets occurred at metamorphosis (26–28 days posthatch). At harvest, all fish, except those fed the two highest n-3HUFA diets, suffered from handling stress (shock syndrome) with increasing severity (25 to 100%) at decreasing dietary n-3HUFA intake. Recovery from shock syndrome was 100% at the 0.63% and 0.87% n-3HUFA diets, 63% at the 0.33% n-3HUFA diet and 0% at the control diet. Post-harvest survival was similar among the four highest dietary n-3HUFA groups (64–73%), whereas the two lowest n-3HUFA groups differed significantly (0 and 37%). Growth promotion was evident with increased dietary n-3HUFA intake as fish fed the highest n-3HUFA diet were twice the size of those fed the control diet (99 vs 52 mg wet-wt). Final fish sizes at the three highest n-3HUFA diets were similar. Given similar culture conditions, a minimum dietary n-3HUFA intake of 1.26% of dry-wt Artermia is recommended to avoid handling stress and promote growth in larval palmetto bass.     

Experimental transmission of Enteromyxum leei Diamant, Lom and Dykova, 1994 in sharpsnout sea bream, Diplodus puntazzo C. and the effect on some innate immune parameters

Enteromyxum leei has been implicated in outbreaks in sharpsnout sea bream, causing a serious economic impact on Mediterranean fish farming. Information about transmission and life cycles of marine myxosporea is limited to only a few fish species. In the present study, direct transmission of the infection was achieved by cohabitation of infected and healthy Diplodus puntazzo C. for the first time. The progress of the infection was monitored by the detection of different developmental stages of the parasite in the posterior intestine. Infected fish with E. leei were firstly observed in day 5 (prevalence of 20%). The total prevalence reached 100% in day 19 and was maintained in this high level until the end of the experimental period (day 55). In addition, the initial intensity of the different parasite stages was low, but on day 26 the intensity of trophozoites was suddenly increased, followed by the increased intensity of sporoblasts and mature spores. Also, the experimental transmission of E. leei in recovered D. puntazzo previously treated for parasite elimination proved to be unsuccessful, suggesting of a possible development of protective immunity against the parasite.     

Gametogenesis of the European flat oyster (Ostrea edulis) and Pacific oyster (Crassostrea gigas) in warm water in Israel

The gametogenic cycle of Ostrea edulis and Crassostrea gigas was studied in the outflow water of fish ponds in Eilat, Israel. Despite relatively high water temperatures (14–28°C) and high salinity (41 ± 2 ppt), both species developed gonads. O. edulis spawned from March to May, when water temperature ranged from 18 to 22°C, with a dormant period during the summer months. C. gigas spawned between May and August, when water temperature ranged between 20 and 26°C, with a dormant period between August and April. Spat never developed in the fish pond system.     

Immunization of grouper, Epinephelus coioides, confers protection against a protozoan parasite, Cryptocaryon irritans

The present study aimed to determine whether protection is conferred by immunization of grouper, Epinephelus coioides, against a protozoan parasite, Cryptocaryon irritans. The immunization of E. coioides was carried out by a low level exposure of fish to live C. irritans theronts from predetermined number of tomonts and by an intraperitoneal injection of a vaccine consisting of formalin-killed C. irritans theronts.

Mucus titers detected by ELISA were significantly higher in fingerling and adult grouper subjected to the low level of exposure to C. irritans theronts at 3-week post-exposure compared to fish that had no previous exposure. In addition, significantly smaller tomonts were produced from adult grouper after three successive exposures than the tomonts produced after a single exposure to the parasite.

In the vaccine-immunization experiment, no mortality was monitored in fish that received high dose vaccine (100 μg/fish), while 40% cumulative mortality and 100% cumulative mortality were recorded in low dose group (10 μg/fish) and control group (PBS-injected), respectively. In the succeeding replicate, the vaccine-immunized group (high dose) had 37.5% cumulative mortality and 100% cumulative mortality for the control. In addition, a total of 1830 tomonts were collected at 5-day post-challenge from the control group while none from the vaccine-immunized group. Significantly fewer trophonts and tomonts were enumerated at 5-day and 7-day post-challenge, respectively, in the vaccine-immunized group than the control.

Results suggest that a protective immunity has been conferred on the immunized grouper as indicated by high antibody titers in the mucus of C. irritans-exposed fish and higher survival and fewer parasites in vaccine-immunized fish than the control groups. The conferred immunity played a major role in preventing or limiting the adhesion, invasion, and development of C. irritans theronts on the skin of the immunized grouper.     

Hydroacoustic in situ target strength of smelt (Osmerus eperlanus (L.))

Dependence of the in situ acoustic target strength (TS) of smelt (Osmerus eperlanus (L.)) on the total fish length (L) was estimated from TS measurements made with a split-beam 120 kHz echo sounder with reference to fish length–frequency distributions in trawl catches. The studies were conducted in the eutrophic and turbid Lake Hiidenvesi in southern Finland. Smelt dominated the catches-in-number with proportions at different sampling stations ranging from 91.8 to 100%. A TS–length equation of the form TS = alog L − b was fitted through the modes in the TS-distributions and in the respective fish length distributions. The estimated model (±parameter standard error) was TS = 23.4 (±2.24)log₁₀ L − 68.7 (±1.82). With a pre-defined slope of 20, the fitted equation was TS = 20 log₁₀ L − 65.9 (±0.23). The estimated day–night difference in TS–length relationship was not significant (t-test, p = 0.37).     

Growth performance of gilthead sea bream Sparus aurata in different osmotic conditions: Implications for osmoregulation and energy metabolism

The influence of three different environmental salinities (seawater, SW: 38 ppt salinity; brackish water, BW: 12 ppt; and low salinity water, LSW: 6 ppt salinity) on the growth, osmoregulation and metabolism of young gilthead sea bream (Sparus aurata L.) was studied over a period of 100 days. 480 inmature fish (20 g mean body weight) were randomly divided into six tanks of 2500 l (80 fish per tank) and maintained under three different salinities (38 ppt, 12 ppt and 6 ppt) in an open system. Every three weeks, 10 fish from each tank were anesthetized, weighed and lenghed. At the end of experiment, 10 fish from each tank were anesthetized, weighed and sampled for plasma, brain, gill and liver. Gill Na⁺, K⁺-ATPase activity, plasma osmolality, ions (sodium and chloride), glucose, lactate, protein and triglyceride, and hepatosomatic index were examined. In addition, levels of glycogen, lactate, ATP and activities of potential regulatory enzymes (hexokinase, pyruvate kinase, glycogen phosphorylase, and glucose 6-phosphate dehydrogenase) were assesed in liver, brain, and gill. BW-acclimated fish showed a better growth with respect to SW- or LSW-acclimated fish (12 > 38 > 6 ppt). The same relationship was observed for weight gain and specific growth rate. Osmoregulatory parameters in plasma (osmolality, Na⁺ and Cl⁻ levels) were similar in SW- and BW-acclimated fish but significantly higher than those of LSW-acclimated fish. Gill Na⁺, K⁺-ATPase activity showed lower values in intermediate salinity (6 > 38 > 12 ppt). No changes were observed in metabolic parameters analyzed in plasma, whereas only minor changes were observed in metabolic parameters of liver, gills and brain that could be correlated with the higher growth rates observed in fish acclimated to BW, which do not allow us to attribute the best growth rate observed at 12 ppt to lower metabolic rates in that salinity.     

Seawater adaptation and growth of post-smolt Atlantic salmon (Salmo salar) of wild and farmed strains

Atlantic salmon smolts (Salmo salar L.) of wild (Namsen) and farmed (AquaGen) strains were transferred to full-strength seawater (33‰) for 0 (initial control group), 0.5, 1, 2, 4, 8, 14, 30, 42 and 60 days at three different temperatures (4.3, 9.4 and 14.3 °C). Freshwater temperature in each tank was adjusted to test conditions 10 days prior to transfer. Physiological adaptation was monitored as changes in plasma growth hormone levels, gill Na⁺,K⁺-ATPase activity, plasma chloride levels and survival in seawater. Overall, smolts from the wild strain were better able to tolerate transfer to seawater than smolts from the hatchery strain. A delay in the osmotic disturbance and a prolonged period of osmotic stress were observed at the low temperature. Circulating GH levels increased transiently in all groups during the first 12–48 h in seawater and long-term GH levels were positively correlated with seawater temperature. Growth rates were influenced by temperature and strain, with the farmed smolts showing a higher growth than the wild smolts. Food Conversion Efficiency (FCE) was higher in smolts from the farmed strain, whereas no differences in daily food consumption were observed. Optimum temperature for FCE was calculated to be 10.5 °C, whereas the optimal temperature for growth in seawater was calculated to be 13.0 °C. We suggest that the observed differences in seawater tolerance, growth and food conversion probably are genetic and may reflect the fact that the hatchery fish have been bred for rapid growth for several generations.     

A study of appetite, digestion and growth in juvenile green turtle (Chelonia mydas L.) fed on artificial diets

One-year-old immature green turtles (500–900 g) were kept at 25°C in recirculating seawater. When deprived of food for 108 h and then offered floating trout pellets, they consumed a satiation meal (S g) described by the equation S = 0.025 W^0.96 where W is wet body weight in g. When food was offered at different deprivation times after satiation, maximum meals were taken after 60–72 h and corresponded to 2.5% body weight. Longer deprivation (108 h) led to reduced intake of 1.9–2% body weight. Gastric emptying was measured using X-radiography or chromic oxide to label the faeces. Complete stomach emptying required 110 ± 11 h S.D. and maximum appetite occurred when the stomach was estimated to be 80–95% empty. The average time for a meal to be voided from the alimentary tract was 176 ± 16 h S.D. Diets containing 40–50% protein and 4.2–5 kcal/gram were assimilated with apparent efficiencies of 76 ± 6% S.D. and 86 ± 6% for energy and protein nitrogen, respectively. A preliminary estimate for an energy budget for these animals based on a 23 day growth period accounted for 90% of the daily food intake as:

100 I39M + 32E

where known daily intake (I calories) is compared with measures or estimates of metabolic rate (M), growth (G) and excreted wastes (E). Although this balance is similar to that of herbivorous fish, the turtle has slower feeding and digestion rates than its teleost counterparts.     

Gut contents of silver carp, Hypophthalmichthys molitrix, and the disruption of a centric diatom, Cyclotella, on passage through the esophagus and intestine

The digestibility of algae by stomachless filter-feeding fish has been debated for decades. Results from gut contents and digestive enzyme analysis suggest poor utilization, while the measurement of food assimilation using radiolabeled isotope techniques indicate reasonable assimilation efficiency. Phytoplankton in the gut contents of the planktivorous filter-feeding silver carp were studied during March–May. The fish were cultured in a large net cage in a shallow, hypertrophic subtropical Chinese lake, Lake Donghu. In terms of biomass, the dominant phytoplankton in the fore-gut contents were Cyclotella (average 77.8%, range 69.7–93.5%) and Cryptomonas (average 9.57%, range 0–20.4%). The Ivlev's electivity index E of silver carp was much higher for Cyclotella (1.54) than for Cryptomonas (0.56). The majority of the phytoplankton found in the intestines of silver carp were 8–20 μm, but they were also able to collect particles as small as 4.5–10 μm, smaller than their filtering net meshes, suggesting that the secretion of mucus may play an important role in collecting such small particles. We conclude that disruption of cell walls is principally by the pharyngeal teeth, explaining previous contradictory conclusions.     

Age, growth, and reproduction of silky sharks, Carcharhinus falciformis, in northeastern Taiwan waters

To examine the age, growth and reproduction of silky sharks, Carcharhinus falciformis, in the waters off northeastern Taiwan, 469 specimens (213 females and 256 males) were collected from August 2000 to January 2002 at the Nanfanao fish market, northeastern Taiwan. The relationship between body weight (W) and total length (TL) for both sexes combined was expressed as: W = 2.92 × 10⁻⁶ TL^3.15 (n = 469, p < 0.01). The relationship between TL and vertebral centrum radius (R) for both sexes combined data was estimated as: TL = 25.979 + 18.197R (n = 250, p < 0.01). Growth bands (including translucent and opaque zones) in precaudal vertebrae formed once a year between December and January and were counted up to 11 and 14 for females and males, respectively. The von Bertalanffy growth function (VBGF) was used to model the observed length at age data. The sexes combined VBGF predicted an asymptotic length (L_∞) = 332.0 cm TL, growth coefficient (k) = 0.0838 year⁻¹, age at zero length (t₀) = −2.761 year (n = 250, p < 0.01). Size at 50% maturity for males was estimated to be 212.5 cm based on the logistic curve, which corresponded to 9.3 years. Females matured at 210–220 cm, which correspond to 9.2–10.2 years. The length at birth was estimated to be 63.5–75.5 cm TL. The number of embryos per litter was 8–10 and sex ratio of embryos was 1:1.     

Vibrio harveyi and other bacterial pathogens in cultured summer flounder, Paralichthys dentatus

A monitoring program for Vibrio harveyi and other potential bacterial pathogens of summer flounder was conducted at two facilities in the Northeast United States. Bacterial samples were collected from larval and juvenile fish and live-feed, and identified using API 20E biochemical profiles and 16S rDNA sequencing. Histopathological examinations were conducted in order to relate histological changes with the presence of potential bacterial pathogens. V. harveyi, Vibrio ichthyoenteri, and Photobacterium damselae subsp. damselae, three known pathogens of flatfish, were isolated from diseased summer flounder. Although a high proportion of juvenile summer flounder showed microscopic signs of disease, the presence of these potential bacterial pathogens in fish was not associated with large-scale mortalities. An outbreak of flounder infectious necrotizing enteritis with 30% cumulative mortality occurred when juveniles were transported to a new facility. Isolates of V. harveyi from the disease outbreak were genetically different from the isolates from the commercial hatchery. V. harveyi isolates from both facilities were pathogenic to summer flounder by intraperitoneal injection. P. damselae subsp. damselae, V. ichthyoenteri, and Vibrio scophthalmi, also found in fish with gross lesions, were not pathogenic to juvenile summer flounder by intraperitoneal injection. Our research shows that several potential bacterial pathogens are associated with morbidity and mortality in summer flounder larvae and juveniles, especially in situations of stress. Increased knowledge about the environmental conditions that lead to disease, as well as the interactions of V. harveyi with other microbial species, could lead to the development of management strategies in summer flounder farms.     

Tank culture of yellowfin tuna, Thunnus albacares: developing a spawning population for research purposes

A land-based culture facility for research on yellowfin tuna, Thunnus albacares, was developed at the Achotines Laboratory in the Republic of Panama. Six concrete tanks, and seawater and life support systems were built to maintain a yellowfin broodstock. On average, 50% of the yellowfin caught survived capture and handling, and approximately 30% became broodstock in Tank 1 (17 m diameter, 6 m depth) or Tank 2 (8.5 m diameter, 3 m depth). Each fish was tagged with a microchip implant tag, then weighed, measured, and injected with oxytetracycline (OTC) prior to stocking. Daily rations of primarily market squid, Loligo opalescens, and Pacific thread herring, Opisthonema spp., were regulated based on the feeding activity and energy requirements of the fish. Feeding activity of the broodstock decreased when the water temperatures decreased, and the fish ate decreasing daily rations and increasing calories with increasing size. Spawning occurred in both tanks within 6–8 months of capture. Spawning first occurred in Tank 1 when 24 females ranged in size from 6 to 16 kg and 65 to 93 cm fork length (FL). Spawning was intermittent during the first 2 months and occurred near daily thereafter. Tank size appeared to affect survival rates, the types of mortalities that occurred, and the growth of the fish. Survival rates after 1 year in captivity were higher, and the fish were larger, on average, in Tank 1 than in Tank 2. Most of the mortalities in Tank 1 were the result of wall strikes, which occurred more frequently after the fish reached their highest density of 0.64 kg m⁻³ and sizes greater than 96 cm FL and 19 kg. Non-linear growth models were fitted to the initial stocking sizes and final sizes of fish that died or were removed from Tank 1 during 1996–1999. Estimated growth rates in length (11–48 cm year⁻¹) for fish between 51 and 150 cm FL decreased with increasing length. Estimated growth rates in weight ranged from 9 to 19 kg year⁻¹ for fish less than 19 kg and 20–23 kg year⁻¹ for fish greater than 19 kg. The results of this work demonstrate that the stable environment of a land-based culture facility may be the preferred system for long-term maintenance of a yellowfin broodstock.     

Nutrients removed by Kappaphycus alvarezii (Rhodophyta, Solieriaceae) in integrated cultivation with fishes in re-circulating water

The potential of the red alga Kappaphycus alvarezii to remove nutrients was tested to treat effluents of Trachinotus carolinus fish cultivation, and the production of carrageenan in this condition was analyzed. Experiments were conducted in four tanks of 8000 L with approximately 1200 fishes of 30 g each integrated with three tanks of 100 L with 700 g of K. alvarezii, as initial biomass per tank. Seawater was re-circulated between tanks with seaweed and with fish. As a control, three tanks with seawater circulating in an open system were utilized. Seawater samples were collected daily for 10 days and concentrations of nitrate, nitrite, ammonium and phosphate were determined in the inflow and outflow water of the tanks. Significant differences between both collecting points were considered as nutrient removal by the seaweed. Growth rates and carrageenan yields were also analyzed in seaweed cultivated in seawater and in effluents. Growth rates of seaweed cultivated in tanks were lower than those obtained in open sea and in laboratory cultivation. Effluents had concentrations of nitrate and nitrite ca. 100 times higher than in the control. Maximum values of nutrient removal on effluents were: nitrate = 18.2%; nitrite = 50.8%; ammonium = 70.5% and phosphate = 26.8%. All plants survived throughout the experimental period, but some developed “ice–ice”, a disease associated with physiological stress. After the experimental period, some plants selected and cultivated in open sea presented higher growth rates in 40 days, indicating nutrient storage. No significant differences between carrageenan yields of K. alvarezii cultivated in seawater and in the effluents were observed. Our results show that K. alvarezii can be utilized as a biofilter for fish cultivation effluents, reducing the eutrophication process and can also be processed for carrageenan production, which provides an additional benefit to the fisheries.