SUMMARY This study is an attempt to gain an integrated understanding of the interactions between temperature, locomotion activity and metabolism in the European sea bass (Dicentrarchus labrax). To our knowledge this study is among the few that have investigated the influence of the seasonal changes in water temperature on swimming performance in fish. Using a Brett-type swim-tunnel respirometer the relationship between oxygen consumption and swimming speed was determined in fish acclimatised to 7, 11, 14, 18, 22, 26 and 30°C. The corresponding maximum swimming speed(Umax), optimal swimming speed (Uopt),active (AMR) and standard (SMR) metabolic rates as well as aerobic metabolic scope (MS) were calculated. Using simple mathematical functions, these parameters were modelled as a function of water temperature and swimming speed. Both SMR and AMR were positively related to water temperature up to 24°C. Above 24°C SMR and AMR levelled off and MS tended to decrease. We found a tight relationship between AMR and Umax and observed that raising the temperature increased AMR and increased swimming ability. However, although fish swam faster at high temperature, the net cost of transport (COTnet) at a given speed was not influence by the elevation of the water temperature. Although Uopt doubled between 7°C and 30°C (from 0.3 to 0.6 m s-1), metabolic rate at Uopt represented a relatively constant fraction of the animal active metabolic rate (40-45%). A proposed model integrates the effects of water temperature on the interaction between metabolism and swimming performance. In particular the controlling effect of temperature on AMR is shown to be the key factor limiting maximal swimming speed of sea bass.

This contribution reviews existing literature and some new own findings on teleost sperm motility and factors controlling it, emphasizing selected marine species. In marine teleosts with external fertilization (halibut, turbot, sea bass, hake, cod and tuna serving as examples), mainly the osmolality controls sperm motility: movement is activated by transfer from the seminal fluid into sea water, representing a large upward step in osmolality. The exception are flatfishes (such as halibut or turbot) where CO 2 is responsible for flagellar immotility in seminal fluid. In all cases, the duration of motility is short and limited to minutes ranges due to partial exhaustion of the ATP energy and to increase of internal ionic concentration as suggested by studies with de-membranated ⁄ ATP reactivated flagellae. In this overview, we compare motility characteristics (percentage of active spermatozoa, velocity, linearity), flagellar waves parameters (wave length and amplitude, number of waves) and energy content (respiration and ATP concentration) within species where these data have been established. All parameters show a rapid decrease after activation; therefore progressive forward movement needed by the sperm to effectively reach the egg surface, is limited to a short initial period following activation. In two species (turbot and sea bass) the rapid decrease of sperm motility is reflected by a corresponding decrease of the fertilizing ability. Exposure to external environments (sea water) at activation also leads to local defects of the sperm flagella posing additional limitations on motility duration. However, minor flagellar damages as well as energetic exhaustion are reversible: after a resting period in a non-swimming solution at the end of the motility period, spermatozoa can be re-activated for a second motility period. From these results and from additional data obtained from de-membranated ⁄ ATP re-activated spermatozoa, a paradigm has been developed which establishes a link between external osmolality (sea water), internal ionic concentration and control of axonemal activity.

After a long period of spermatogenesis (several weeks to months), marine fish spermatozoa are delivered at male spawning in seawater (SW) at the same time as ova. In some fish species, as the ova micropyle closes quickly after release, these minute unicells, the spermatozoa, have to accomplish their task of reaching the micropyle within a very brief period (several seconds to minutes), for delivery of the haploid male genetic information to the ova. To achieve this goal, their high-performance motile equipment, the flagellum, must fully activate immediately on contact with the SW and then propel the sperm cell at an unusually high initial velocity. The cost of such 'hyperactivity' is a very rapid consumption of intracellular ATP that outstrips the supply. The spermatozoa become rapidly exhausted because mitochondria cannot compensate for this very fast flagellar energy consumption. Therefore, any spermatozoon ends up with two possibilities: either becoming exhausted and immotile or reaching the egg micropyle within its very short period of forward motility (in the range of tens of seconds) before micropyle closure in relation to both contact of SWand cortical reaction. The aim of the present review is to present step by step the successive events occurring in marine fish spermatozoa from activation until their full arrest of motility. The present knowledge of activation mechanisms is summarized, as well as a description of the motility parameters characterizing the motility period. As a complement, in vitro results on axonemal motility obtained after demembranation of flagella bring further understanding. The description of the sperm energetic content (ATP and other high energy compounds) and its evolution during the swimming period is also discussed. A general model aiming to explain all the successive cellular events occurring immediately after the activation is presented. This model is proposed as a guideline for understanding the events governing the sperm lifespan in the marine fish species that reproduce through external fertilization.

European hake (Merluccius merluccius) were caught alive in the northern Bay of Biscay in June/July 2002, using a codend specially designed to avoid crushing fish and also to retain water while hauling the trawl. In all, 1307 fish were tagged with anchor T-bar tags, injected with tetracycline, then released. The length range of the tagged and marked fish varied from 13 to 58 cm, and the modal size was 28 cm, at which length they were assumed to be 2–3 years old. A mean survival rate of 68.2% was estimated. Mortality was mainly caused by stress of capture and physical damage, and depended on the size of the catch and the depth of trawling. By the end of March 2003, 32 fish and three tags had been returned to the laboratory (a 2.7% return rate), the recapture data indicating that the probability of survival after release does not depend on catch depth or depth at location of release. For combined sexes, the somatic growth rate was estimated at 0.033 ± 0.019 cm day−1 (n=15). Males and females did not differ significantly in somatic growth rate, which were, respectively, 0.028±0.018 (n=6) and 0.033 ± 0.012 cm day−1 (n=6). This pilot experiment represents the first recorded mass tagging of European hake, which is known to be a very fragile species. The preliminary results indicate that it would be possible to carry out a large-scale tagging experiment of the European stocks in order to improve assessment and subsequent management decisions.

The objective of this paper is to characterize some of the sperm motility parameters in European hake (Merluccius merluccius), which is considered to be a species with aquaculture potential. The total ATP, ADP and AMP concentrations were determined using high-performance liquid chromatography on hake sperm samples collected during the winter-early spring in the Bay of Biscay (France) (n = 22) and on hake sperm samples collected during the summer-early autumn in waters off Western Norway (n = 5). The Adenylate Energy Charge (AEC) was deduced from these data. Computer Assisted Sperm Analysis (CASA) was used to measure a series of parameters characterizing the motility and the sperm swimming performances. Changes in salinity of the swimming medium affected all the measured motility parameters. The sperm velocity and the straightness of the movement were at maximum when sperm was activated with 100% filtrated sea water (100 SW) but decreased sharply later. When sperm was activated in filtrated sea water (50% diluted with distilled water: 50 SW) the values of these parameters increased (with a lower percentage of active cells) during the first 2.5 min and thereafter decreased slowly. In 50 SW, the initial velocity was lowered but the swimming period lasting 4.5 times longer than in 100 SW (but with a lower percentage of actively swimming cells). Initial sperm motility (percentage of swimming cells) in 100 SW was affected by sperm storage duration. Undiluted sperm could be stored at 4°C for 5 days and still show 13 ± 7% motility; the velocity and straightness of the movement were at maximum at the earliest period of measurement (0.5-1 day of storage) and then decreased gradually to reach their minima after 4 days of storage. Further, both the AEC and ATP content decreased with storage time, with the AEC decreasing from 0.78 ± 0.07 (mean ± SD) at stripping time to 0.20 ± 0.09 after 2 days of storage. Over the same period ATP content decreased from 85 ± 80 to 5 ± 4 nanomoles 10 )9 spermatozoa, these data presenting a high variability.

The objective of this study was to complement published hake (Merlucius merlucius) sperm motility characteristics by obtention of data more specifically dependent on flagella: beat frequency, flagellar wave amplitude, wave length, number of waves along the length of the flagellum and curvature of the wave. The changes of these parameters are described in relation to the time period elapsed since activation are described using high resolution video images of flagella while quantifying the progression during the swimming period from the activity initiation to full cessation of movement several minutes later. The main characteristic of the swimming period of hake spermatozoa shows a progressive decrease of forward displacement resulting from a cumulative decrease of several activity descriptors: the beat frequency, the percentage of motile cells, the wave amplitude, the number of flagellar waves and the linearity and shape of flagella. All these parameters decrease within 7-8 min after activation in a converging fashion, which leads to full immotility. Measurements of motility parameters at different temperatures also bring additional information about energetic needs for motility of hake sperm. The wave shape of the flagella showed that: (i) the contact of flagella with sea water, not only triggers immediately its motility, but also provokes rapidly local osmotic damages such as blebs which partly impair the correct wave propagation from head vicinity to flagellar tip; (ii) fully developed waves become more and more restricted to the proximal flagellum while the tip becomes devoid of any wave; (iii) the wave amplitude decreases as a function of time during the motility period. The combination of these factors contributes to motility limitations (<100 s) during which the spermatozoa are efficiently able to reach the egg for fertilization. Temperature effects (5-32°C) on these performance characteristics were also studied; this allows to conclude that the amount of energy needed to sustain motility becomes rapidly limiting. A quantitative evaluation of all these sperm motility features leads to a better definition of the conditions for controlled propagation of this species. The motility parameters are also useful as quantitative descriptors of sperm swimming ability in situations such as: (i) varying salinity (osmolality) of the medium (swimming solution) and (ii) activating motility in seminal fluid at various dilution ratios in sea water. All together, our results lead to a better understanding of hake sperm biological features, including its sperm flagella behaviour.

Dissections of mature and non-mature European hake males and females (N=142) collected in waters off the western coast of Norway and in the Bay of Biscay (France) in 2004-2006 demonstrate for the first time that this gadoid species contains drumming muscles. There were differences in drumming muscles weight with body length, sex and maturity stage. This study shows that the difference between females and males is primarily manifested during the spawning season, seen both in the French and Norwegian samples. For the mature females the drumming muscles dry weight increases only slightly, if at all, with increase in total length. For mature males there is a corresponding rapid increase. There does not seem to be any consistent difference between the average dry weight of the drumming muscles in immature male and immature and mature female hake of the same length, tested on the Norwegian samples.Our results suggest that male hake, like the males of other gadoids studied, may produce sounds in the context of spawning.

Due to stock declines there is a growing interest in farming of European hake Merluccius merluccius. Thus, knowledge of its sperm biology is of importance not only for purposes of broodstock management, but also for the development of sperm preservation techniques. Hake sperm were collected from mature males caught during the summer-early autumn waters off western Norway and during the winter-early spring in the Bay of Biscay (France). Sperm quality characteristics were assessed after storage at 4 °C for 25 ± 14 h. Average (± SD) values for Norwegian and French samples respectively were (i) sperm volume: 3.9 ± 4.0 and 2.6 ± 4.0 ml; (ii) spermatozoa concentration: 6.6 ± 3.2 and 13.9 ± 5.1 × 10 9 spermatozoa/ml; (iii) spermatocrit: 80.2 ± 3.3 and 81.8 ± 10.7%; and (iv) total number of spermatozoa: 23.5 ± 30.0 and 35.1 ± 36.2 × 10 9. Average osmolality and pH (± SD) of French samples were 349 ± 28 mOsmol/kg and 7.6 ± 0.1, respectively. Activation by transfer into full sea water (100 SW) or 10% ovarian fluid in sea water (10 OF) occurred synchronously for virtually all spermatozoa and the percent sperm motile decreased with post activation time. When transferred into 50% sea water diluted with distilled water (50 SW) only a few spermatozoa were activated initially but subsequently reached a maximum percentage of motility followed by a decline. Hake sperm motility declined rapidly to 50% of motility 70 s after activation with 100 SW. Sperm were motile for longer when activated with 50 SW (1570 ± 295 s) or 10 OF (718 ± 71 s) compared to 100 SW (317 ± 121 s). Undiluted hake sperm stored at 4 °C up to 10 days retained 10% motility when activated with 100 SW. When cryopreserved, the motility recovery index of the cells at thawing ranged from 0 to 76.4%. These results describe for the first time the sperm traits of European hake following successful cryopreservation, and also show the importance of activation medium on sperm motility.