Evaluation of head-only and head-to-tail electrical stunning of farmed eels (Anguilla anguilla, L.) for the development of a humane slaughter method

The overall objective was to evaluate the suitability of electronarcosis as a stunning method for farmed eels. In the first experiment the minimum electrical current needed to induce a general epileptiform insult by head‐only stunning was assessed. The individual eels (n = 40) with a live weight of 700–800 g were fixed in a specially designed re‐strainer. The EEG (electroencephalogram) and ECG (electrocardiogram) recordings, observation of behaviour and responses to pain stimuli were used to assess unconsciousness, insensibility and cardiac function. The applied current of 150, 200 or 250 V, 50 Hz, AC was delivered via scissor‐model stunning tongs for approximately 1 s. A general epileptiform insult was observed in 31 eels for which a successful EEG recording was obtained, using 255 ± 4 V, 545 ± 32 mA, for 1.2 ± 0.2 s. The general epileptiform insult as measured on the EEG was characterized by a tonic/clonic phase and an exhaustion phase. The behaviour showed one phase: tonic cramps alternated by clonic ones. The heart rate was 22 ± 8 beats min^?1 (n = 23) prior to stunning. After stunning the ECG revealed fibrillation. In the second experiment the behaviour of seven individual eels able to move freely in water was observed after head‐only stunning (250 V). Two phases were distinguished. Limited tonic and clonic cramps combined with backward swimming were followed by heavy clonic cramps combined with unco‐ordinated movements such as jumping out of the water. A distinct exhaustion phase was not observed in all animals. In the third experiment a head‐to‐tail electrical method was examined in 15 eels for rendering the eels unconscious and insensitive prior to slaughter. They were stunned by applying 253 V for 3 s followed by 50 V for 5 min. In the fourth experiment nine eels were head‐only stunned with 260 V for 1 s immediately followed by 50 V for 5 min applied from head to tail. Results obtained in these two experiments were similar. After stunning no brain activity and no responses to pain stimuli on the EEG were observed and the ECG showed ventricular extra systolae. It was observed that it might take 60 ± 25 min or longer for a complete recovery. It can be concluded that for effective electrical stunning of eels with a weight of 700–800 g an average current of 545 ± 32 mA (at approximate 250 V, 50 Hz AC) is needed. In this case, within a confidence level of 95% at least 91% of the eels are effectively stunned (n = 31). Therefore, it is recommended to increase the minimum current for an effective stun in practice to 600 mA. Further research is needed to determine the conditions to induce permanent unconsciousness and insensibility of the eels to protect the animals at slaughter.     

Evaluation of Electrical Stunning of Atlantic Cod (Gadus morhua) and Turbot (Psetta maxima) in Seawater

The aim of this study was to assess electrical stunning of Atlantic cod and turbot in seawater to develop a protocol for the process of stunning and killing. An induced general epileptiform insult (unconscious) had a duration of 40 ± 27 s (n =14) in cod (2.6 ± 0.5 kg) and 34 ± 18 s (n = 19) in turbot (520 ± 65 g). Seven cod and 3 turbot displayed a physical reaction, and 11 turbot registered an electroencephalogram (EEG) response to pain stimuli administered 30 s post-stun. The heart rate was 32 ± 6 beats/min in cod and 25 ± 7 beats/min in turbot prior to stunning. Post-stunning, the electrocardiogram (ECG) revealed fibrillation and reduced activity post-stun. EEG, ECG recordings, and behavioral observations indicate that when a bipolar square wave current was applied with a frequency of 133 Hz and 43% duty cycle side to side (turbot) and at 170 Hz and 33% duty cycle (cod) head to tail, both species were stunned in seawater at current densities of 3.2 A/dm² and 2.5 A/dm², respectively. For turbot, a 5 s exposure to electricity followed by chilling in ice water for 15 min is sufficient to prevent recovery. For cod, a killing method needs to be established.     

Electrical stunning followed by decapitation or chilling of African catfish (Clarias gariepinus): assessment of behavioural and neural parameters and product quality

The objective of this study was to assess neural and behavioural responses in farmed African catfish (Clarias gariepinus) upon electrical stunning in combination with decapitation or chilling. To assess the possibility of scaling up one or both experimental methods, two trials were performed in an experimental setting. The product quality of the collected samples was compared with the currently applied industrial method: live chilling. After electrical stunning in combination with decapitation, the fish showed spikes alternated with theta and delta waves on the EEG, followed by minimal brain activity after 20±10 s. The same traces on the EEGs were observed after electrical stunning in combination with chilling. Here, minimal brain activity occurred after 22±11 s. Within a confidence level of 95%, the percentage of African catfish that was effectively stunned after administration of an electrical current of 1.5 A dm⁻², 300 V (50 Hz a.c.), followed by decapitation or chilling was above 91%. The analysis yield and evolution of liquid loss showed significant (P<0.05) differences among the batches, which could be explained by the stunning method used. The course values of the pH in the different batches were significantly (P<0.05) dependent on the stunning method, sex and location (visceral or skin side). It is concluded that African catfish can be stunned effectively using electrical stunning in a water tank, followed by decapitation or chilling in ice. Dutch commercial processors prefer to combine electrical stunning with chilling in flake ice in a rotating tumbler, as the outer slime layer is then removed, which facilitates further processing.     

Stunning of farmed African catfish (Clarias gariepinus) using a captive needle pistol; assessment of welfare aspects

The objective of this study was to assess whether high‐pressure injection of air into the brain of African catfish (Clarias gariepinus) could render the animal unconscious and insensible immediately and permanently. In the study, 48 African catfish with a live weight of 900–1900 g were restrained and equipped with EEG and ECG electrodes and then stunned. The catfish were stunned mechanically using a captive needle pistol. The pressure to shoot the needle was 8 bar and that to inject the air was 3 bar for 1.5 s. The catfish behaviour was observed during and after stunning. τ and δ waves and spikes, which precede a stoppage in brain activity as measured on the EEG, were used as indices for the measurement of immediate induction of unconsciousness and insensibility In 23 of 42 fish, an iso‐electric line was observed after an average of 13.4 s, while in the remaining fish the τ and δ waves and spikes remained on the EEG during the recording period. In all cases, the ECG showed an irregular heart rate with fibrillation and extra systolae. Moreover, the configuration showed ischaemia. Before the captive needle stunning, free‐swimming fish (n=7) explored the tank for an average of 21±12 s before lying down at the bottom. After stunning, they showed clonic uncoordinated swimming movements. The movements stopped after an average of 38±50 s. In another group (n=7) that was stunned and subsequently placed in ice water, clonic cramps were observed in two out of seven animals. When taking into account the number of animals with a reliable EEG (n=42) and using 95% confidence intervals, it was concluded that at least 93% of the catfish were effectively stunned using a correctly positioned captive needle pistol. Furthermore, it is recommended to immobilize the stunned fish by chilling, as the post‐stun clonic cramps may hinder gutting and filleting.     

Evaluation of electrical stunning of sea bass (Dicentrarchus labrax) in seawater and killing by chilling: welfare aspects, product quality and possibilities for implementation

The objective was to assess neural, behavioural responses and product quality in farmed sea bass (Dicentrarchus labrax) upon electrical stunning in seawater. The electrical sinusoidal 50 Hz or pulse square wave alternating 133 Hz current induced a general epileptiform insult with a current of 3.3±0.2 or 3 A dm^?2, respectively, for 1 s head to tail in seawater. The total duration of the insult was 48±34 and 23±11 s. After stunning, the electro‐cardiogram revealed fibrillation and ceased or showed malfunction. Product quality was assessed in a group electrically stunned, followed by chilling in ice water and the controls were only chilled in ice water. The pH of the fillets was 0.1–0.2 lower (P<0.01) when stunned electrically at days 1, 2, 8 and 10 postmortem, where the colour did not differ. The percentage of sea bass effectively stunned using an electrical sinusoidal or pulse square wave current was above 85% within a confidence level of 95%. A combination of electrical stunning for 10 s, followed by chilling in seawater with ice flakes resulted in the death of all fish. The former method is recommended to be adapted for implementation in practice.     

Is humane slaughter of fish possible for industry?

Abstract The objective was to evaluate industrial and research slaughter methods for Atlantic salmon (Salmo salar), gilt‐head seabream (Sparus auratus) and eel (Anguilla anguilla) with respect to welfare and quality. As a general term of reference, an optimal slaughter method should render fish unconscious until death without avoidable excitement, pain or suffering prior to killing. For Atlantic salmon, commercial slaughter methods (carbon dioxide stunning followed by gill cutting, and gill cutting alone) are not in conformity with the general term of reference, as the fish are not rendered unconscious immediately and possibly experience stress. Evaluation of automated percussive stunning remained unconclusive. More research should enable us to ascertain whether loss of consciousness is instantaneous. Electrical stunning can be humane if applied properly. However, because flesh of electrostunned fish was characterized by occasional bloodspots, optimization of the electrical parameters is required. Prototypes for percussive and electrical stunning of salmon have been recently developed. This implies that humane slaughter of salmon is feasible for industry. For gilt‐head seabream, neither aphyxia in air nor transfer of the fish to an ice slurry were considered to be humane: the methods did not induce immediate brain dysfunction and vigorous attempts to escape occurred. Percussive and electrical stunning can be in conformity with the general term of reference. However, conditions for stunning whole batches of seabream have not been established. Quality of the fish slaughtered by percussive stunning was similar to that obtained by the industrial method, i.e. immersion in an ice slurry. Further work is required to establish optimal stunning conditions and to develop prototypes. For eel, desliming in a salt‐bath followed by evisceration, electrical stunning performed under the conditions prescribed by the German legislation, and live chilling and freezing were not considered to be humane. In contrast, it was established that a 10–20 kg batch of eels in fresh water could be rendered unconscious immediately and until death by applying electricity in combination with nitrogen gas. The conditions used were 0.64 A dm^?2 for 1 s, followed by 0.17 A dm^?2 combined with nitrogen flushing for 5 min. A preliminary assessment of flesh quality suggests that it may be improved by application of the latter method, compared with the salt bath. The results clearly indicated that humane slaughter of eels is possible in practice.     

Welfare aspects of live chilling and freezing of farmed eel (Anguilla anguilla L.): neurological and behavioural assessment

The overall objective of the study was to evaluate a slaughter method of eels, which consisted of chilling until their body temperature was <5 °C for stunning, and subsequently placing them in cold brine at −18 °C for 15 min for killing. Three distinct experiments and a control were performed.

Firstly, 19 eels with an average live weight of 758±44 g were restrained and equipped with EEG, ECG electrodes and a temperature sensor inside the body. Then, they were placed in the ice water. Indices for the induction of unconsciousness and insensibility were the appearance of theta and delta waves and no response on pain stimuli, which disappeared at a body temperature of 8.0±2.1 °C after 12±5 min in 15 eels. The responses to pain stimuli did not disappear in three eels. Within a confidence level of 95%, the percentage of eels that is not effectively stunned during the procedure in ice water of <5 °C was at least 5%. The heart rate decreased from 24±10 beats/min (n=14) to 7±4 (n=11) and became irregular during cooling down. When placed in the brine water of −18 °C, the EEG showed rapid and extreme depolarisation of the membranes, which started after 27±17 s (n=18). The ECG showed fluttering of the heart in all eels. None of the eels recovered after this procedure.

For 10 eels with an average live weight of 128±27 g, it was observed that the body temperature decreased from 17.1±0.6 to 4.0±0.5 °C in the ice water. After 15 min in the brine water of −16.1±2.2 °C, the body temperature decreased to −3.1±2.3 °C.

Finally, three groups of seven eels and eight single eels were placed in ice water of −0.0±0.1 °C. The observation of unrestrained eels revealed four phases. Animals were (1) swimming around in the water, (2) attempting to escape from the ice water, (3) pressing their nose to the wall or corner while showing clonic muscle cramps, and finally (4) breathing only, while all other muscle activity was totally suppressed. Afterwards, they were transferred to cold brine at −18 °C, and none of the eels recovered.

The eight control eels, which were transferred to water at 18 °C, swam around, except for one that was lying in an S-shape position at the bottom. After 570 and 605 s, two eels tried to escape from the box.

The obtained results show that the eels, which were transferred from water at 18 °C to ice water, might be stressed, a specific behaviour and an irregular heart rate were observed. From an animal welfare point of view, it is therefore not recommended to stun eels by live chilling. Moreover, at least 5% of the eels will not be stunned at a body temperature of <5 °C. Placing eels in brine water of −18 °C is an effective method to kill the eels before slaughter. However, it cannot be recommended to place conscious eels in cold brine water, because it takes more than 27 s before unconsciousness may be induced.     

Electrical stunning of farmed Atlantic cod Gadus morhua L.: a comparison of an industrial and experimental method

An industrial and experimental electrical method for stunning farmed Atlantic cod in air and seawater (SW), respectively, were compared. The impacts of sedation with AQUI‐S^? and exercise to exhaustion before electrical stunning were also assessed to monitor the possible depletion of rested muscle energy levels by electrical stunning. Stress (blood glucose, haematocrit, muscle pH, muscle excitability, high‐energy phosphates and rigor mortis) and flesh quality (fillet texture, colour, liquid leakage (LL), gaping, residual blood and K‐value) were assessed. For the industrial stunning method, an average of 41 V, 0.2 A dc was applied to individual cod for 18–27 s. For the SW method, a bipolar square wave current (170 Hz, 33% duty cycle) was applied for 5 s. After stunning, recovery was prevented by exsanguination in chilled SW. There were no differences (P>0.05) between the two stunning methods except for a higher ultimate fillet pH for cod stunned in air 8 days postmortem. Exercise before stunning depleted muscle energy levels at slaughter, increased LL and fillets had redder and darker flesh after storage on ice for 8 days. Electrical stunning (in air) of AQUI‐S^?‐treated fish partly depleted muscle energy levels (pH 7.3, ATP 18.7 μmol g^?1, PCr 70.1 μmol g^?1). However, flesh quality was not affected. Unless pre‐rigor filleting is the chosen processing strategy, electrical stunning of cod seems to be a promising stunning method.     

Growth differences of Japanese eels Anguilla japonica between fresh and brackish water habitats in relation to annual food consumption in the Kojima Bay‐Asahi River system,Japan

To learn about the relationships between feeding and growth of temperate eels in freshwater and brackish water habitats, we analysed 533 yellow‐phase Japanese eels Anguilla japonica collected in both types of habitats in southeastern Japan. Because male eels were very rare in each habitat (FW, N = 1; BW, N = 20), characteristics of female eels were compared between the different habitats. Annual food consumption was evaluated with the consideration of instantaneous food consumption and annual activity period. Stomach fullness index (stomach content weight/body weight) was used as an indicator of instantaneous food consumption. The ratios of number of months with eel catch to those when eel sampling was conducted were used as an indicator of activity period. Female yellow eels tended to be older and slower growing in fresh water (N = 78; age, mean ± SD = 7.9 ± 2.4 years; growth rate, 59.8 ± 14.0 mm year^?1) than in brackish water (N = 229; age, 5.5 ± 1.8 years; growth rate, 90.1 ± 24.4 mm year^?1). Irrespective of sex, yellow eels in brackish water had a higher stomach fullness index and a greater ratio of months with eel catches, indicating greater annual food consumption by brackish water eels. These results indicate that greater annual food consumption contributes to the greater growth rates of Japanese eels in brackish water habitats.     

Evaluation of Quality Parameters in the Meat of Pintado Amazônico,Female Pseudoplatystoma fasciatum × Male Leiarius marmoratus,Subjected to Different Slaughter Methods

The objective of this study was to check the effect of different stunning methods on the meat quality of Pintado Amazônico, female Pseudoplatystoma fasciatum × male Leiarius marmoratus, stored on ice for 18 d. A total of 90 specimens (2.5 ± 0.45 kg and 58.21 ± 6.20 cm) were divided into three groups and subjected to stunning by: water saturated with CO₂, hypothermia on ice, or asphyxia in air. Subsequently, samples were removed to be analyzed after (times) 0, 2, 4, 6, 8, 10, 12, 14, 16, and 18 d. At the established times, aerobic mesophilic heterotrophic microorganisms and aerobic psychrotrophic heterotrophic microorganisms were quantified, and the pH, total volatile base nitrogen, color (L*a*b*), and sensory traits were analyzed. The stunning method significantly affected (P < 0.05) the meat quality. The group stunned on ice showed the best results compared with the other treatments.     

Site fidelity,diel and seasonal activities of yellow‐phase Japanese eels (Anguilla japonica) in a freshwater habitat as inferred from acoustic telemetry

Japanese eels (Anguilla japonica) perform large‐scale oceanic migrations between their spawning ground and growth habitats in continental waters during life history. However, between these migrations, they spend most of time in growth habitats such as rivers. To investigate the diel and seasonal activities, homing behaviour and home range of yellow‐phase Japanese eels in the lower reach of the Tone River, we tracked them throughout a year using a fine‐scale positioning system (VPS) based on acoustic telemetry. The tracked eels were generally nocturnal, but not exclusively. They were mainly mobile from spring to autumn, with little or no activity observed during winter. A transport‐release experiment showed that most eels returned to their original capture area within 13 days after release. The eels had very small home ranges (mean ± SD = 0.085 ± 0.068 km²), core areas (0.014 ± 0.014 km²) and linear home ranges (744 ± 268 m). They also tended to be distributed on one particular side of the river (right or left bank) and in one particular shore type (revetment or vegetation), rarely moving from one to the other. This study provides evidence for nocturnal, dormancy, homing behaviours, limited habitat use and small home range size in Japanese eels. The eels clearly showed strong fidelity to a “familiar” site, which contrasts with the long distances travelled during upstream and downstream migration phases in the river, and during spawning migrations in the ocean.     

Variation in the demographic characteristics of yellow‐phase Japanese eels in different habitats of the Hamana Lake system,Japan

Abstract – Yellow‐phase Japanese eels (Anguilla japonica) were investigated in the Hamana Lake system, Japan, from 2003 to 2004 to understand how their demographic attributes vary within the lake system. A total of 779 yellow eels were collected during sampling in two inlet rivers and two brackish/saltwater lakes within the lake system. Female eels predominated, constituting 84% of the 75 sex‐determined eels in the river, and 50% of the 151 sex‐determined eels in the lakes. Total lengths (TL) of all eels examined ranged from 54.2 to 715.0 mm (mean = 320.4 ± 145.4 SD). In the inlet river, the TL of eels showed a significant positive relation with the distance from the river mouth. The estimated relative abundances of eels ranged from 0 to 1.8 eels·m^?2 effort (mean: 0.3 ± 0.41) in the river and was negatively correlated with the distance from the river mouth. This suggested that larger eels might tend to be distributed at lower abundances in upstream reaches of the river. Mean age of yellow eels determined by their otolith annuli was younger in the lake (N = 117, 3.3 ± 1.4 years) than in the river (N = 214, 4.3 ± 1.7 years). Growth rate was higher in the lake than in the river at age 1–2 years (131.9 and 104.4 mm·year^?1, respectively). The results of this study suggest that, although Japanese eels can adapt to various types of environments, significant differences can occur in population structures and growth patterns among habitats.     

Effects of salinity and temperature on the growth and survival of New Zealand shortfin,Anguilla australis,and longfin,A. dieffenbachii,glass eels

The ideal water conditions for maximizing the performance of the nursery culture of glass eels harvested from the wild for aquaculture need to be determined for the New Zealand shortfin (Anguilla australis) and longfin (Anguilla dieffenbachii) eels. This study determined the survival and growth of glass eels reared under different temperature and salinity conditions in the laboratory. The growth and survival of shortfin and longfin glass eels reared in salt water (35‰) maintained at 25 °C was examined over 84 days from capture. The mean specific growth rate (SGR) was higher in shortfin [2.30±0.29% body weight (b.w.) day^?1] than longfin glass eels (1.52±0.06% b.w. day^?1), and survival was also higher in shortfin (76.0±4.16%) than for longfin glass eels (28.7±6.36%). A second experiment identified the effect of salinity (0, 17.5‰ and 35‰) and temperature (17.5 and 26.5 °C) on the acclimation, growth performance and survival of shortfin and longfin glass eels over a period of 84 days from capture. There was no incidence of mortality for either shortfin or longfin glass eels reared across all salinity treatments (0‰, 17.5‰ and 35‰) at 26.5 °C, while survival of shortfin and longfin glass eels reared at 17.5 °C was the highest in 17.5‰, followed by 35‰ and 0‰ treatments. Both temperature and salinity affected the SGR of shortfin glass eels, with the highest SGR observed for shortfin glass eels reared in 0‰ water maintained at 26.5 °C. In longfin glass eels, salinity alone had an effect on the SGR, with the highest SGR observed in glass eels reared in 0‰ water regardless of the water temperature (17.5 and 26.5 °C). In addition, the adaptability of glass eels to salinity was evaluated from the development and the physiological responses of gill chloride cell (CC) morphology. The number and size of CCs increased significantly with increasing salinity in both shortfin and longfin eels.     

Direct and indirect impacts of pumping station operation on downstream migration of critically endangered European eel

Downstream passage of European eel Anguilla anguilla (L.) in catchments with pump(s) for water level management is a major concern. Catchment‐wide acoustic telemetry revealed silver eels quickly migrated downstream through unobstructed reaches (n = 12; mean ± SD = 17.9 ± 1.9 km/day). Fourteen of 17 acoustic‐tagged eels detected at the pumping station (82.1%) retreated back upstream and ten (58.8%) passed through pumps after delays (9.5 ± 11.0 days). Multi‐beam sonar imaging across the intake screen (55‐mm gaps) revealed that peaks in migration occurred during the nights preceding the new moon but 76.7% retreated back upstream. All passive integrated transponder (PIT)‐tagged eels recaptured (n = 56) downstream of a large (2.23‐m diameter) mixed flow pump survived but 96.5% had minor injuries, reduced physical condition and/or abnormal behaviour. By contrast, 64.7% of PIT‐tagged eels recaptured (n = 17) downstream of a small (0.8‐m diameter) axial flow pump died. No acoustic‐tagged eels that passed through the small axial flow pump (n = 10) performed onward migration at sea, unlike “control” eels released downstream (n = 11). This evidence may help develop effective remediation measures, such as operational changes, to maximise escapement of catadromous eel species at pumping stations.     

Effects of salinity on standard metabolic rate and critical oxygen tension in the giant freshwater prawn (Macrobrachium rosenbergii)

The extensively farmed giant freshwater shrimp, Macrobrachium rosenbergii, can survive salinities up to 26 g L^?1, but the commercially important grow‐out occurs exclusively in freshwater areas. Recent studies suggest the shrimp equally capable of growing in brackish as fresh water and a better understanding of how this species responds to changing salinity could significantly impact freshwater prawn farming in deltas and coastal areas. Here, the effect of salinity (0 and 15 g L^?1) on standard metabolic rate (SMR) and critical oxygen tension (P_crit) was measured in adult M. rosenbergii using intermittent closed respirometry. SMR was 79.8 ± 3.1 and 72.7 ± 2.9 μmol kg^?1 min^?1 in fresh and brackish water, respectively, with no significant difference between the two salinities (P = 0.122). During hypoxia M. rosenbergii maintained oxygen uptake down to a P_crit of 26.3 ± 1.4 mmHg in fresh and 27.2 ± 2.0 mmHg in brackish water (P = 0.682), showing that salinity had no overall effect on oxygen conductance in the animals. These findings are in agreement with recent growth studies and provide further evidence that grow‐out phase could be accomplished in brackish water areas. Thus, the predicted intrusions of brackish water in tropical deltas as a consequence of future global warming may not impact this important production.     

Recovery from transportation by road of farmed European eel (Anguilla anguilla)

The objective of this study was to assess the effects of transportation of marketable eel (0.15 kg) in the Netherlands with respect to welfare. Eels (Anguilla anguilla) were obtained from a commercial farm and acclimatized for 7 weeks at the laboratory. Fish were transported according to regular commercial procedures. The animals were placed in water‐filled transport tanks on the trailer. Fish density increased from 72 kg m^?3 (husbandry) to 206 kg m^?3 (fasting) and was further increased to 270–290 kg m^?3 during transport. Fish transport lasted 3 h after which the eels were returned to laboratory recirculation systems to measure parameters indicative of stress load, i.e. mortality, plasma cortisol, lactate and non‐esterified fatty acids (NEFA) as well as gill morphology. Samples were taken at 0, 6, 24, 48 and 72 h after transport in transported fish and non‐transported counterparts (controls). Transportation affected water quality within known tolerable limits. No mortality during or after transport was observed. After 6 h, plasma cortisol levels had returned to baseline. However, energy metabolism had increased suggesting that transportation of eels resulted in an increased energy demand that lasted for at least 72 h in the fasted animals. Thus, it is conceivable that exposure to adverse conditions, prior to stunning/killing, in a slaughterhouse may result in allostatic overload in eel.     

A comparative epizootiologic study of the two fish‐pathogenic serovars of Vibrio vulnificus biotype 2

Vibrio vulnificus biotype 2 is subdivided into two main serovars, serovar E, able to infect fish and humans, and serovar A, only virulent for fish. Serovar E emerged in 1976 as the causative agent of a haemorrhagic septicaemia (warm‐water vibriosis) affecting eels cultured in brackish water. Serovar A emerged in 2000 in freshwater‐cultured eels vaccinated against serovar E, causing warm‐water vibriosis with fish showing a haemorrhagic intestine as the main differential sign. The aim of the present work was to compare the disease caused by both serovars in terms of transmission routes, portals of entry and host range. Results of bath, patch‐contact and oral‐anal challenges demonstrated that both serovars spread through water and infect healthy eels, serovar A entering mainly by the anus and serovar E by the gills. The course of the disease under laboratory conditions was similar for both serovars in terms of transmission and dependence of degree of virulence on water parameters (temperature and salinity). However, the decrease in degree of virulence in fresh water was significantly greater in serovar E than in serovar A. Finally, both serovars proved pathogenic for tilapia, sea bass and rainbow trout, but not for sea bream, with significant differences in degree of virulence only in rainbow trout. In conclusion, serovar A seems to represent a new antigenic form of V. vulnificus biotype 2 with an unusual portal of entry and is better adapted to fresh water than serovar E.     

Cryopreservation of Atlantic cod (Gadus morhua) sperm in large‐volume straws: applications for commercial production and gene banking

In our study, we used a full factorial analysis of variance design to examine the effects of diluent [Mounib's sucrose‐based diluent+hen's egg yolk (EY) and Hanks' balanced salt solution (HBSS)+EY], freezing rate (?2.5, ?5.0 and ?7.5 °C min^?1) and thawing rate (2.5, 5.0 and 7.5 °C min^?1) on motility and velocity of Atlantic cod sperm cryopreserved in 2.5 mL cryogenic straws. We found that post‐thaw sperm performance was strongly influenced by the presence of higher‐order interactions of the factors we tested. For all models broken down by diluent, the 2.5 °C min^?1 thawing rate had the lowest sperm motility recovery index. Mounib's sucrose‐based diluent+EY had the highest motility recovery index at all thawing rates. Mean per cent motility for fresh sperm (87.7±2.9%) was not significantly different than of sperm cryopreserved using Mounib's sucrose‐based diluent+EY, frozen at ?2.5 °C min^?1 and thawed at 5.0 °C min^?1 (77.1±2.9%). For Mounib's sucrose‐based diluent+EY, velocity was significantly higher with sperm thawed at 7.5 °C min^?1, than sperm thawed at 2.5 °C min^?1, while thawing rate had no effect for HBSS+EY. Our findings have implications for cod mariculture and aiding in conservation efforts for a dominant marine fish species.     

Comparison of the effects of six stunning/killing procedures on flesh quality of sea bass (Dicentrarchus labrax,Linnaeus 1758) and evaluation of clove oil anaesthesia followed by chilling on ice/water slurry for potential implementation in aquaculture

The effects of different stunning/killing procedures on flesh quality of European sea bass were investigated: (1) anaesthesia with clove oil, (2) anaesthesia with 2‐phenoxyethanol, (3) percussive stunning, (4) immersion in ice/water slurry, (5) chilling on ice and (6) anaesthesia with clove oil followed by immersion in ice/water slurry. Muscle pH values were significantly lower in sea bass anaesthetized or stunned by a blow to the head compared with fish immersed in ice/water slurry, chilled on ice or immersed in ice/water slurry after clove oil anaesthesia. Lightness was highest in sea bass anaesthetized by 2‐phenoxyethanol or percussively stunned and lowest in ice‐chilled fish. Redness and yellowness were highest in fish chilled on ice and lowest in fish anaesthetized with clove oil. Liquid loss, fat loss and shear values were not significantly different among the procedures. In general, lipid oxidation values during refrigerated or frozen storage did not significantly differ among treatment groups. Combination of clove oil anaesthesia followed by chilling on ice/water slurry appears to improve both flesh quality and welfare of sea bass, although the implementation of further studies is warranted to consolidate this finding.     

Production and storage of sperm from the black sea bass Centropristis striata L

Black sea bass Centropristis striata L. are protogynous hermaphrodites that develop and spawn as females before changing sex to male. Since all fish eventually become males, determining the relationship between sperm production, sperm quality and seasonal changes in plasma levels of testosterone (T) and 11‐ketotestosterone (11‐KT) could be useful for identifying appropriate males to maintain as broodstock. Milt and blood samples were collected three times during an 8‐week spawning season. Milt volume (3.5±0.76 mL kg^?1), sperm density (3.2 × 10⁸± 0.31 cells mL^?1), sperm production [11 × 10⁸±3.4 cells kg^?1 body weight (BW)] and sperm motility (80±0.6%) were at their highest during the first sampling interval and coincided with the highest 11‐KT levels (1.0± 0.11 ng mL^?1). All of the sperm indices decreased to their lowest levels during the final 3 weeks of the study. Sperm viability was highly correlated (adjusted R²=0.84) with sperm motility. Sperm cryopreserved in modified Mounib's extender (MME) had the highest post‐thaw motility compared with two other extenders. Post‐thaw motility of sperm cryopreserved in MME was not different from fresh after 90 days of storage. There was no difference in fertilization rates between fresh (69±2.4%) and post‐thaw (67±4.1%) sperm samples taken from the same male or among males. These results demonstrate that the quality of black sea bass spermatozoa is higher earlier in the spawning season and that acceptable post‐thaw fertilization rates can be obtained from cryopreserved sperm.