Sensory Evaluation of the Off-Flavors Geosmin and 2-Methylisoborneol (MIB) in Farm-Raised Catfish

Individuals responsible for tasting catfish in quality control settings sometimes report sensations of sensory adaptation, sensory fatigue and flavor carry-over between samples containing earthy/muddy off-flavors due to geosmin and/or 2-methylisoborneol (MIB). If these experiences are real, sensory evaluation of flavor quality may not be as straightforward as many fish processors and researchers would wish. To determine the influence of these compounds on the sensory evaluation of fish, a series of controlled experiments was conducted. These studies indicate that trained panelists were unable to recognize and evaluate, with precision, the intensity of off-flavor compounds such as geosmin and MIB. However, panelists were precise in evaluating other natural off-flavors, such as fishy. In evaluating two MIB samples in succession, the second sample was perceived as having a lower MIB intensity. The addition of off-flavor compounds to catfish caused the panelists to perceive the desirable flavor chickeny to be lower in intensity, indicating masking. Based on these experiments, a simplified off-flavor quality rating system, consistent with available data, is proposed.     

The effects of salinity on the survival, growth and haemolymph osmolality of early juvenile blue swimmer crabs, Portunus pelagicus

The blue swimmer crab, Portunus pelagicus, is an emerging aquaculture species in the Indo-Pacific. Two experiments were performed to determine the effects of salinity on survival, growth and haemolymph osmolality of early juvenile P. pelagicus crabs. The salinities tested for the first experiment were 10, 15, 25 and 40 ppt, and for the second experiment 5, 20, 30, 35 and 45 ppt. Each salinity experiment was triplicated, with each replicate consisting of 10 stage 4 juveniles. Each experiment lasted 45 days. Mortalities and incidence of “molt death syndrome” were recorded daily, while the intermolt period, carapace length, carapace width and wet weight were measured at each molt. At the end of the experiments the haemolymph osmolality and dry weights were measured.

Results demonstrate that salinity significantly affects both the survival and growth of early P. pelagicus juveniles. Mortality was significantly higher (p < 0.01) for juveniles cultured at salinities ≤ 15 ppt and at 45 ppt. At a salinity of 5 ppt a complete mortality occurred on day 20. In all salinity treatments, the majority of mortalities were due to “molt death syndrome”. In experiment 1, immediate effects of salinity on growth and development were seen at 10 ppt as the intermolt period was significantly longer (p < 0.01) and the mean carapace size increase was significantly less (p < 0.01) at the first molt compared to the other treatments. Meanwhile, the specific growth rates (carapace length, width and wet weight) were significantly lower (p < 0.05) at high salinities (≥ 40 ppt) due to longer intermolt periods and significantly lower (p < 0.05) carapace size or wet weight increases.

The haemolymph osmolality exhibited a positive linear relationship with the culture medium with an isosmotic point of 1106 mOsm/kg, equal to a salinity of approximately 38 ppt. Based on the osmolality graph, high metabolic cost for osmoregulation due to increased hyper- and hypo-osmotic stress appeared to cause lower survival and specific growth rates of the crabs. The results demonstrate that a salinity range of 20–35 ppt is suitable for the culture of early juvenile P. pelagicus.     

Evaluation of the impact of nitrate-nitrogen levels in recirculating aquaculture systems on concentrations of the off-flavor compounds geosmin and 2-methylisoborneol in water and rainbow trout (Oncorhynchus mykiss)

Aquatic animals raised in recirculating aquaculture systems (RAS) can develop preharvest “off-flavors” such as “earthy” or “musty” which are caused by the bioaccumulation of the odorous compounds geosmin or 2-methylisoborneol (MIB), respectively, in their flesh. Tainted aquatic products cause large economic losses to producers due to the inability to market them. Certain species of actinomycetes, a group of filamentous bacteria, have been attributed as the main sources of geosmin and MIB in RAS. Previous studies have demonstrated that certain nutritional factors can stimulate or inhibit bacterial biomass and geosmin production by certain actinomycetes. In the current study, the effects of two nitrate-nitrogen (NO₃^--N) levels (20–40 mg/L and 80–100 mg/L) on geosmin and MIB levels in culture water and the flesh of rainbow trout (Oncorhynchus mykiss) raised in RAS were monitored. Water and fish tissue samples were collected over an approximately nine-week period from six RAS, three replicates each of low and high NO₃^--N, and analyzed for geosmin concentrations using solid phase microextraction–gas chromatography–mass spectrometry. Results indicated no significant difference in geosmin concentrations in water or fish flesh between the low and high NO₃^--N RAS. Therefore, higher NO₃^--N levels that may occur in RAS will not adversely or beneficially impact geosmin-related off-flavor problems.     

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.     

Geosmin and Off-Flavor in Nile Tilapia (Oreochromis niloticus)

Abstract

The concentration of geosmin (1,10-trans-dimethyl-trans-9-decaol) in fresh tilapia were analyzed by applying high-vacuum distillation, extraction and gas chromatography techniques. The distribution of geosmin in the fish was determined by examining tissues from various parts of tilapia, along with the rates of geosmin absorption and purging. Analysis showed that when geosmin was added to fish flesh at concentrations ranging from 2.8 to 390 μg per kg of flesh, the rate of recovery was 51 to 89%. Sensory evaluation detected muddy flavor in freshwater tilapia with geosmin content ranging from 7.55 to 9.85 μg/kg of fish flesh. However, panelists failed to detect muddy flavor in brackish water tilapia that contained geosmin only in the range of 1.5 to 2.6 μg/kg. Among various tissues of the fish, the intestines contained the highest geosmin concentration, and appeared in descending order in the abdominal, skin and muscle tissues. After holding for 2 h in water with 5 and 50 μg/l geosmin, tilapia absorbed 17.6 and 42.2 μg/kg geosmin in the flesh, respectively. Holding the fish over longer periods in water with higher geosmin content resulted in a greater amount of absorption. In contrast, transferring muddy-flavored fish to static clean water purged geosmin from the fish but it required at least 16 days to eliminate the muddy flavor.     

Effects of Preservation Methods On Geosmin Content and Off-Flavor in Nile Tilapia (Oreochromis niloticus)

Abstract

In this study, the masking or reduction of off-flavor in tilapia due to various preservation methods such as salting, drying, frying, smoking, microwave heating, marinating and fermentation with carbohydrate mixture (som fak preparation) was investigated by subjecting the processed tilapia to sensory evaluation and analyzing the concentrations of geosmin (1,10-trans -dimethyl-trans -9-decaol) in the processed samples. Dry salting or brining muddy-flavored fish and then drying either by hot air at 50°C or sun-drying resulted to only a slight reduction in the geosmin content of the product. Deep-frying reduced the muddy flavor intensity and geosmin content in salted-dried tilapia. Pretreatment of tilapia fillets with acidified brine before smoking reduced geosmin content and masked the muddy flavor in the smoked product. Microwave cooking of fresh muddy-flavored tilapia showed no effect on its geosmin content nor its off-flavor. Marinating tilapia in acetic acid solution resulted in decreased muddy flavor, and longer marinating period led to lower geosmin content in the product. The geosmin content of som fak made from muddy-flavor and non-muddy-flavor tilapia differed significantly, although sensory evaluation yielded no significant differences between the two types of som fak, and the taste panelists preferred the product fermented for 3 days.     

Impact of starvation and handling stress on rigor development and quality of Atlantic salmon (Salmon salar L)

Farmed Atlantic salmon (Salmo salar L) were fed to satiation or starved for 35 d, and subjected to acute pre-slaughter stress or careful handling before slaughtering. Rigor development, post-mortem energy metabolism and quality variations were analysed during 72 h cold storage. The body weight averaged 2.9 and 3.5 kg of the fed and starved salmon, respectively. The starved salmon had lower condition factor (0.99 vs. 1.11) and higher slaughter yield (92.6 vs. 87.5%), but the fat content was similar (11.8–12.9%) in the fed and starved salmon. The initial glycogen level was lower in the starved salmon (21 vs. 29 μmol g^− 1), whereas initial ATP levels (6.4–6.6 μmol g^− 1) were not affected by starvation. Pre-slaughter stress accelerated rigor development, stimulated lactate formation through post-mortem glycolysis, and increased the breakdown of ATP and CP. The effect of stress on rigor development was less pronounced in starved salmon, and a significant interaction was observed between nutritional status and stress. Rigor development was closely correlated to ATP breakdown, especially in fillets. Acute stress accelerated flesh softening during ice storage. After 72 h storage, the fillets of the starved salmon exposed to careful slaughter handling had the significantly firmest texture, whereas the most intense fillet colour was observed in the starved/stressed salmon. Liquid leakage from the muscle during cold storage was lower in the starved salmon, but pre-slaughter stress had no significant impact. It is concluded that a starvation period of five weeks can improve the resistant to acute stress prior to slaughtering of Atlantic salmon that in turn hampers rigor development.     

Inter-specific variation of the mitochondrial r16S gene among silversides, “Peces Blancos”, (Atherinopsidae: Menidiinae) and its utilization for species identification

Analysis of 570 nucleotide sequences of the mitochondrial r16S gene of four species of silversides “peces blancos” (Chirostoma sp.) and of two euryhaline related species Membras martinica and Menidia menidia indicated variation that aided in development of a DNA-based method for species identification. The nucleotide sequence data showed levels of inter-generic genetic variation that ranged from 3.3–7.9% and from 1.06 to 1.6% at the inter-specific level. The variation observed allowed the identification of 18 diagnostic restriction enzymes that in any combination of two or three could aid in the diagnosis of species of “peces blancos”; six of these enzymes could also discriminate between the genera Membras and Menidia; the enzymes AseI, CviAII and FatI discriminate the genus Membras from Chirostoma; and the combination of any of the enzymes BsaAI, CviAII, FatI or HpyCH4IV discriminate Menidia from the “peces blancos”. PCR-RFLP analysis of amplified mitochondrial r16S fragments of 90 un-identified “peces blancos” from Lake Patzcuaro utilizing the restriction enzymes HpyCH41V, TaqI, BsmF1 and BbvI allowed us to diagnose unambiguously cultivated Chirostoma estor and C. humboldtianum in juvenile and adult stages. The use of a non-invasive method to PCR-amplify specific DNA markers from live brood stocks suggests use of this tool for identification of endangered or threatened species.     

Effect of peracetic acid on levels of geosmin, 2-methylisoborneol,and their potential producers in a recirculating aquaculture system for rearing rainbow trout (Oncorhynchus mykiss)

In recirculating aquaculture systems (RAS)s, off-flavors and odors, mainly caused by geosmin (GSM) and 2-methylisoborneol (MIB), can accumulate in the flesh of fish from RAS water, reducing the profitability of production. In this study, peracetic acid (PAA) was applied in three application intervals to pump sumps of rainbow trout (Oncorhynchus mykiss) reared in RAS. Using a real-time polymerase chain reaction (qPCR), the potential off-flavor producers were quantified using geoA and MIB synthase genes. Streptomyces was identified as the major GSM producer, and biofilters showed the highest number of potential off-flavor producers. Concentrations of GSM and MIB were analyzed in the circulating water and in the lateral part of the fish fillet. In water, concentrations up to 51 ng L⁻¹ (GSM) and 60.3 ng L⁻¹ (MIB) were found, while in the fillet, these were up to 9.8 ng g⁻¹ (GSM) and 10.2 ng g⁻¹ (MIB), decreasing with increasing number of PAA applications. PAA applications reduced the levels of off-flavor compounds, although this was insufficient to fully prevent the accumulation of GSM and MIB.     

Pharmacokinetics and the active metabolite of enrofloxacin in Chinese mitten-handed crab (Eriocheir sinensis)

The pharmacokinetics and active metabolite of enrofloxacin were estimated after single intramuscular administration (10.0 or 20.0 mg/kg body weight) to the Chinese mitten-handed crab (Eriocheir sinensis) in fresh water at 25.0 ± 1.0 °C. Levels of enrofloxacin and its metabolite ciprofloxacin in the main tissues (hemolymph, hepatopancreas, muscle, ovary and spermary) were simultaneously detected by HPLC. Enrofloxacin concentration–time profiles for the hemolymph in both tests were described by a two-compartment open model with first-order absorption. Distribution half-time (T_1/2), elimination half-time (T_1/2β), body clearance (CL/F), mean residence time (MRT_0–∞), area under the concentration–time curve from 0 to ∞ h (AUC_0–∞) and apparent volume of distribution (Vd/F), which derived from the pharmacokinetic model, were 0.427 h, 21.3 h, 0.133 l/h/kg, 60.0 h, 96.9 μg/ml/h and 4.08 l/kg, respectively, at a dose of 10.0 mg/kg body weight, and 0.216 h, 12.3 h, 0.189 l/h/kg, 85.8 h, 187 μg/ml/h and 3.35 l/kg, respectively, at a dose of 20.0 mg/kg body weight. Similarities were found between the hemolymph concentration–time curves of the two tests; for example, instant absorption process followed by the distribution phrase, and a second absorption peak at 6 h post-treatment. After intramuscular administration of 10.0 mg/kg body weight, absorption of enrofloxacin was observed in the main edible tissues (hepatopancreas, muscle, ovary and spermary), and the drug residue was the highest in the hepatopancreas, where the ‘drug sink’ phenomenon occurred. Comparative pharmacokinetics showed fast absorption, broad distribution and fast elimination of enrofloxacin in E. sinensis after intramuscular dosing. Regarding ciprofloxacin, the main active metabolite of enrofloxacin, though relatively low levels were detected in all the main tissues of the crab, its kinetics in the hemolymph in the two tests were not described by a one- or two-compartment open model.     

Extenders and cryoprotectants for cooling and freezing of piracanjuba (Brycon orbignyanus) semen, an endangered Brazilian teleost fish

The aim of this study was to develop a protocol for semen storage of piracanjuba (Brycon orbignyanus) by both cool storage at 4 °C and cryopreservation at − 196 °C. Semen was diluted in some fish semen extenders (Exp. 1) or in extenders combined with the antibiotic gentamycin sulfate (Exp. 2) and stored at 4 °C. Sperm motility was estimated every 24 h. Then, the effects of egg yolk (0 and 5%), cryoprotectants (dimethyl sulphoxide — DMSO, methanol, and methylglycol) and extenders (NaCl 154 mM, BTS™ Minitub and M III™ Minitub) on semen cryopreservation were evaluated (Exp. 3). Semen was added to each of eighteen cryosolutions (2 yolk concentrations × 3 cryoprotectants × 3 extenders), aspirated into 0.5-mL straws, frozen in nitrogen vapor (Taylor-Wharton, CP 300, “dry shipper”) and stored at − 196 °C. Sperm motility was evaluated after thawing at 60 °C-water bath for 8 s. The three cryosolutions that produced the highest post-thaw sperm motility were used again to freeze semen. Post-thaw semen quality was then evaluated under three tests: sperm motility, the percentage of live spermatozoa and hatching rate (Exp. 4). Piracanjuba semen diluted (1:10 total volume) in NaCl 200 mM or in Saad solution (NaCl 200 mM, Tris 30 mM) maintained motility above 35% for as long as 7 days, at 4 °C. Motility of only 7% was observed on undiluted semen after 3 days at 4 °C. There was neither beneficial nor detrimental effect of gentamycin on sperm motility at 250 μg/mL. Egg yolk addition to the cryosolution was beneficial in samples cryopreserved in NaCl 154 mM and in M III™, but detrimental for samples cryopreserved in BTS™. Methylglycol was the most effective cryoprotector compared to DMSO and methanol. Motility and percentage of live spermatozoa were similar among semen cryopreserved in NaCl–yolk, M III™–yolk and BTS™, all containing 10% methylglycol, but lower than fresh control. Hatching rates of eggs fertilized with sperm cryopreserved in NaCl–yolk or BTS™ were higher than for eggs fertilized with sperm cryopreserved in M III™–yolk, but lower than control fertilizations. The semen cryopreservation protocols developed here will be used to set up a gene bank for endangered piracanjuba populations.     

Identification of conditions underlying production of geosmin and 2-methylisoborneol in a recirculating system

Geosmin and 2-methylisoborneol (MIB) are semi-volatile terpenoid compounds produced as secondary metabolites by benthic and planktonic cyanobacteria, several genera of fungi, and various actinomycetes. These off-flavor compounds pose a heavy economic burden in the aquaculture industry rendering fish unmarketable unless purified by purging with large quantities of clean water. In the present study, the presence of off-flavor compounds was examined in a recirculating aquaculture system (RAS) for tilapia culture. In this zero-discharge system, where water from the fish basins is recirculated through parallel aerobic (drum filter and a trickling filter) and anaerobic treatment loops (sedimentation/digestion basin), concentrations of geosmin and, in particular, MIB were highest in the aerobic treatment loop. Lowest concentrations were detected in the anaerobic treatment loop. This latter finding pointed toward a possible reduction of these compounds in this basin. Two bacterial strains of the streptomycetes family were isolated from the aerobic, organic-rich, drum filter and the nitrifying trickling filter. In vitro tests with these isolates, closely related to Streptomyces roseoflavus and Streptomyces thermocarboxydus, revealed that MIB production exceeded geosmin production under all conditions tested and was significantly higher under aerobic than under anoxic conditions. Under the latter conditions, with nitrate as an electron donor, the S. roseoflavus-like isolate was capable of denitrification. Based on the results obtained in this study, it was concluded that aerobic, organic-rich conditions stimulate the growth of actinomycetes and subsequent production of geosmin and MIB in the system. The observed reduction of these compounds in the anaerobic water treatment component may serve in designing treatment steps aimed at alleviating the problem of geosmin and MIB accumulation in recirculating systems.     

Catches of swimming crabs using fish mince in “teabags” compared to conventional fish baits in collapsible pots

Fishing trials were conducted in a pond using collapsible pots containing fish mince inside a “teabag” or cut fish baits. Scomber japonicus was minced (100 g) and packed in a teabag or cut in half; non-baited pots served as controls. Baits were placed inside box-shaped pots (2.3 cm mesh size) with two slit entrances. Trials consisted of 100-pot hauls/bait type using a 1-day soaking time. The crab catches of Charybdis japonica and Portunus pelagicus from “fish” and “mince teabag” baits were not significantly different, but differences were found for total catch and damselfish Chromis notata. Trials using 80 g of mince and 20 g of sugar as the “teabag” bait lowered the catch of C. japonica, Thalamita sima and Thalamita prymna but not P. pelagicus when compared with fish bait. Results indicate that the mince in teabags is as suitable as fish bait, and that adding substances to the mince might be used to make selective crab baits.     

Development of phytoplankton communities and common off-flavors in a biofloc technology system used for the culture of channel catfish (Ictalurus punctatus)

The use of biofloc technology production systems continues to increase in the aquaculture industry worldwide. Recent research demonstrated that outdoor biofloc systems can be used to produce high yields of channel catfish (Ictalurus punctatus). However, studies have not yet been performed to determine the development and composition of phytoplankton communities and related off-flavor problems in these biofloc production systems. In this study, water samples were collected biweekly from May to November and channel catfish samples were collected during harvest in November from nine 18.6 m² biofloc culture tanks. Water and fillet samples were analyzed for levels of the common off-flavor compounds geosmin and 2-methylisoborneol (MIB). The development and composition of phytoplankton communities in each culture tank was also monitored. In addition, water and biofloc samples were evaluated to assess the microbial sources of geosmin and MIB within the culture tanks. Phytoplankton (including algae and cyanobacteria attached to bioflocs) biomass, as determined by concentrations of chlorophyll a in the water, gradually increased in all tanks over time. Phytoplankton communities that developed in the culture tanks were dominated by fast-growing, unicellular and small colonial types of green algae (chlorophytes) and diatoms (bacillariophytes) and slower growing, small colonial types of cyanobacteria (cyanophytes). A positive correlation (p < 0.05) between cumulative feed addition and chlorophyll a concentration was found. Although geosmin and MIB were present in the culture water of each tank during most of the study, levels were typically low and only one tank yielded catfish with geosmin and MIB in their flesh at levels high enough to be designated as off-flavor. A positive correlation (p < 0.05) between cumulative feed addition and MIB concentrations in the water of culture tanks indicates a greater potential for MIB-related off-flavor problems at high feed application rates. The microbial sources responsible for production of geosmin and MIB in the culture tanks remain unknown.     

Efficacy, tissue distribution, and residue depletion of oxytetracycline in WS-RLP infected California red abalone Haliotis rufescens

Oxytetracycline (OTC) was recently demonstrated to be an effective therapeutant for California red abalone (Haliotis rufescens) infected with the agent of withering syndrome (WS), a Rickettsia-like prokaryote (WS-RLP). This study examined the influence of temperature on the efficacy and pharmacokinetics of oral OTC treatments on WS-RLP infected red abalone, H. rufescens. Medication was administered in an artificial diet containing 1.85% active OTC at a rate of 103.4 mg/kg abalone for 10, 20 and 30 days at both 13.4 °C and 17.3 °C. Drug residue concentrations, WS-RLP burden and WS-associated pathological changes within foot muscle and digestive gland were measured 3, 17, 23, 42, 63, 81, 102, 122, and 160 days after OTC treatment. In all samples the highest concentrations of OTC were detected on the first sample day. Furthermore, all sampling periods showed significantly less OTC in foot muscle samples relative to corresponding digestive gland samples (p < 0.001). While drug depuration occurred rapidly from foot muscle, unusually high concentrations and long depuration times were measured within the digestive gland. The pharmacokinetic trends in both the foot muscle and digestive gland at 13.4 °C were adequately described by a one-compartment model, while the digestive gland depuration models within the 17.3 °C treatments conformed to a two-compartment model. At both temperatures treatment duration gave rise to significant differences in OTC accumulation and depletion (p < 0.001). Despite this, terminal elimination rate constants were similar. Estimated half-lives were similar among all treatments for digestive gland (23.8–27.5 d) and all treatments for the foot muscle at 13.4 °C (10.7–13.4 d; calculated only for the 13.4 °C treatments). At both temperatures all three durations of drug treatment led to significant reductions in WS-RLP prevalence, foot muscle atrophy, and mortality. WS-RLP re-challenge trials initiated 44, 88, and 122 days following the 10-day treatment at 17.3 °C showed reduced susceptibility to re-infection for up to 88 days after treatment, suggesting that digestive gland OTC residues confer long-term resistance to pathogen. Atomic absorption spectroscopy conducted on both foot muscle and digestive gland found significantly higher concentrations of iron, zinc, and manganese in the digestive gland, offering an explanation behind the underlying mechanism of OTC retention for extended periods within this tissue.     

Disrupted sexual cycles in female grass carp (Ctenopharyngodon idella) raised in tropical conditions

In the Ivory Coast, grass carp (Ctenopharyngodon idella) have been integrated in the tilapia-based polyculture in order to increase pond productivity. In the tropical conditions prevailing there, female cycles seemed disrupted. We describe oogenesis in these conditions using histological observations, monitoring of individual cycles with intraovarian biopsies, endocrinal monitoring (development of a specific enzyme-linked immunosorbent assay (ELISA) for vitellogenin (Vg), measurement of plasma oestradiol-17β (E2) and testosterone (T)) and comparison with grass carp raised in Poland. We noted that oogenesis was blocked in all females at the migrating germinal vesicle stage, precluding ovulation or spawning without artificial induction. High rates of atypical post-vitellogenic oocytes (translucent, not filled with yolk granules) were observed in many females. Female individual cycles also displayed atypical features: cycles were sometimes (10% of females) blocked at the beginning of vitellogenesis (BV), for females displaying abnormally low E2 (0.5 ng/ml) and Vg (30 μg/ml) levels compared to “normal” females (1.4 ng/ml and 223 μg/ml, respectively). The duration of the cycles was highly variable among females (a few days to several weeks). Sexual cycles were unsynchronised (all the ovarian stages could be found for all seasons) and the number of females at the end of vitellogenesis (EV) was low (<40% most of the year). These characteristics raise problems for artificial induction of spawning in small-scale hatcheries: a large stock of broodfish is required as is regular checking of female broodstock with intraovarian biopsies to select responsive females.     

Comparing denitrification rates and carbon sources in commercial scale upflow denitrification biological filters in aquaculture

Aerobic biological filtration systems employing nitrifying bacteria to remediate excess ammonia and nitrite concentrations are common components of recirculating aquaculture systems (RAS). However, significant water exchange may still be necessary to reduce nitrate concentrations to acceptable levels unless denitrification systems are included in the RAS design. This study evaluated the design of a full scale denitrification reactor in a commercial culture RAS application. Four carbon sources were evaluated including methanol, acetic acid, molasses and Cerelose™, a hydrolyzed starch, to determine their applicability under commercial culture conditions and to determine if any of these carbon sources encouraged the production of two common “off-flavor” compounds, 2-methyisoborneol (MIB) or geosmin. The denitrification design consisted of a 1.89 m³ covered conical bottom polyethylene tank containing 1.0 m³ media through which water up-flowed at a rate of 10 lpm. A commercial aquaculture system housing 6 metric tonnes of Siberian sturgeon was used to generate nitrate through nitrification in a moving bed biological filter. All four carbon sources were able to effectively reduce nitrate to near zero concentrations from influent concentrations ranging from 11 to 57 mg/l NO₃–N, and the maximum daily denitrification rate was 670–680 g nitrogen removed/m³ media/day, regardless of the carbon source. Although nitrite production was not a problem once the reactors achieved a constant effluent nitrate, ammonia production was a significant problem for units fed molasses and to a less extent Cerelose™. Maximum measured ammonia concentrations in the reactor effluents for methanol, vinegar, Cerelose™ and molasses were 1.62 ± 0.10, 2.83 ± 0.17, 4.55 ± 0.45 and 5.25 ± 1.26 mg/l NH₃–N, respectively. Turbidity production was significantly increased in reactors fed molasses and to a less extent Cerelose™. Concentrations of geosmin and MIB were not significantly increased in any of the denitrification reactors, regardless of carbon source. Because of its very low cost compared to the other sources tested, molasses may be an attractive carbon source for denitrification if issues of ammonia production, turbidity and foaming can be resolved.     

Use of avoidance response by rainbow trout to carbon dioxide for fish self-transfer between tanks

Convenient, economical, and reduced labor fish harvest and transfer systems are required to realize operating cost savings that can be achieved with the use of much larger and deeper circular culture tanks. To achieve these goals, we developed a new technology for transferring fish based on their avoidance behavior to elevated concentrations of dissolved carbon dioxide (CO₂). We observed this behavioral response during controlled, replicated experiments that showed dissolved CO₂ concentrations of 60–120 mg/L induced rainbow trout (Oncorhynchus mykiss) to swim out of their 11 m³ “growout” tank, through a transfer pipe carrying a flow with ≤23 mg/L dissolved CO₂, into a second 11 m³ “harvest” tank. The research was conducted using separate groups of rainbow trout held at commercially relevant densities (40–60 kg/m³). The average weight of fish ranged from 0.15 to 1.3 kg during the various trials. In all trials that used a constant flow of low CO₂ water (≤23 mg/L) entering the growout tank from the harvest tank, approximately 80–90% of the fish swam from the growout tank, through the transfer pipe, and into the harvest tank after the CO₂ concentration in the growout tank had exceeded 60 mg/L. The fish that remained in the growout tank stayed within the area of relatively low CO₂ water at the entrance of the transfer pipe. However, the rate of fish transfer from the growout tank to the harvest tank was more than doubled when the diameter of the transfer pipe was increased from 203 to 406 mm. To consistently achieve fish transfer efficiencies of 99%, water flow rate through the fish transfer pipe had to be reduced to 10–20% of the original flow just before the conclusion of each trial. Reducing the flow of relatively low CO₂ water near the end of each fish transfer event, restricted the zone of relatively low CO₂ water about the entrance of the fish transfer pipe, and provided the stimulus for all but a few remaining fish to swim out of the growout tank. Results indicate that the CO₂ avoidance technique can provide a convenient, efficient, more economical, and reduced labor approach for fish transfer, especially in applications using large and well mixed circular culture tanks.     

Variation in Channel Catfish Ictalurus punctatus Flavor Quality and its Quality Control Implications

Abstract Producers may capture two to three channel catfish Ictalurus punctatus L. for flavor analysis from ponds scheduled for harvest. If off-flavors are not present in several consecutive fish samples, the population may be considered acceptable for harvest. However, instrumental analysis of the muddy/earthy off-flavor metabolites 2-methylisoborneol (1-R-exo-1,2,7,7-tetramethyl-bicyclo-[2,2,1]-heptan-2-ol) (MIB) and geosmin (lα, 10β-dimethyl-9α-decalol) concentrations in approximately 80 catfish from each of three commercial ponds found both acceptable (on-flavor) and unacceptable (off-flavor) individuals within a single pond (i.e., mixed-flavor populations). Ascertaining the frequency of mixed-flavor populations by instrumentally determining the off-flavor metabolite concentration in muscle tissues from a large number of fish sampled from many ponds at several locations over time is not currently feasible. However, analysis of 12,725 commercial processor flavor assessments collected in 1994 and 1995 indicated 120 instances of individual ponds yielding off-flavor fish followed by on-flavor fish in one day or less. Reports indicate that fish require approximately four days to depurate off-flavors fully, suggesting that a proportion of these rapidly changing flavor assessments may reflect the stochastic selection of fish from mixed-flavor populations rather than a complete and rapid conversion of the flavor quality of entire populations. Factors that contribute to the occurrence of mixed-flavor-populations have not been identified fully. However, increased proportion of fat in catfish fillet tissues has been reported to be correlated with a greater retention of MIB by fish. In this investigation, fish fat contents ranged from 4.45% to 30.45%, and were positively correlated (P < 0.0001) with MIB concentrations. Additionally, the spatial distributions of algal populations within certain commercial catfish ponds were not uniform, and the sensory analysis of the flavor intensity of MIB and geosmin in catfish was more variable than the assessment of the intensity of chickeny and nutty flavors. Probability analysis indicated that in certain mixed-flavor populations there was a 10 to 25% probability of a shipment of fish being rejected due to the random collection of only on-flavor fish prior to harvest followed by the post harvest capture of at least one off-flavor fish. In addition, a chance of no off-flavor fish being sampled from a population containing a proportion of unacceptable fish was indicated. At current sample sizes, replacing sensory analysis with instrumental analysis would not completely avoid problems associated with sampling mixed-flavor-populations. Until effective means to reduce/avoid off-flavor metabolite accumulations in fish are widely available, careful attention to proper sensory evaluation protocols, an enhanced attention to pond conditions that affect flavor quality, the production of leaner more uniform populations of fish, and optimizations of sampling strategies offer the most practical near-term augmentations of current practice.