Sperm motility and seminal fluid composition in the burbot, Lota lota

Sperm motility and composition of the seminal fluid in Lota lota were investigated. Fives after motility initiation, 88.2 ± 12.4% of the spermatozoa were motile, their mean average path swimming velocity was 61.6 ± 16.3 μm s^?1 and their principal swimming type the linear motion (77.4 ± 20.9%). In distilled water the rate of motile spermatozoa decreased to 0% in 40s. In 25–50 mosmol kg^?1 electrolyte (NaCl) or non-electrolyte (glucose, sucrose) solutions, motility was prolonged for 10s and these solutions can therefore increase the efficiency of artificial fertilization when used for sperm motility activation. When semen was diluted in electrolyte or non-electrolyte solutions with osmolalities higher than 50 mosmol kg^?1, sperm motility rates and swimming velocities decreased, and at osmolalities of 400 mosmol kg^?1 motility was completely suppressed. In the seminal fluid with an osmolality of 290.08 ± 45.22 mosmol kg^?1, sodium levels of 139.86 ± 23.79 mmol × 1^?1, potassium levels of 11.59 ± 2.45 mmol × 1^?1 and calcium levels of 0.20 ± 0.08 mmol × 1^?1, sperm motility was inhibited. Under in vitro conditions, artificial saline solutions resembling the seminal plasma composition and 400 mosmol kg^?1 NaCl or glucose solutions were useful as motility inhibiting solutions for predilution of semen. Sperm motility was not affected by pH 7.5–9.0, but at pH 6 the motility rate and the swimming velocity were reduced; seminal fluid pH was 8.47 ± 0.02. Therefore buffering of the artificial saline solutions can provide more stabile conditions for semen during storage and activation. Temperature optimum of semen was between 2 and 5°C. At higher temperatures semen became spontaneously motile. Therefore, controlled temperature conditions are an important factor for handling of semen. The qualitative, organical composition of seminal fluid was similar as in other fresh water teleosts.     

Sperm motility of the marine teleosts Boops boops, Diplodus sargus, Mullus barbatus and Trachurus mediterraneus

The spermatozoa of Boops boops, Diplodus sargus, Mullus barbatus, and Trachurus mediterraneus were motile in sea water, and in electrolyte solutions (NaCl) and non-electrolyte solutions (glucose) with an osmolality of 600–1000 mosmol kg^?1. Their mean motility rate 10 s after initiation was about 80%, while about 10% of the motile spermatozoa moved non-linearly, 45% linearly, and 45% circularly. The average path swimming velocity was significantly higher in M. barbatus (about 90 μm s^?1) than in the other species (70 μm s^?1). The number of motile spermatozoa decreased to 0% within 50 s after initiation of motility in T. mediterraneus, within 90 s in M. barbatus . In B. boops and D. sargus about 90% of the spermatozoa stopped movement during the first 90 s of the motility period, while the rest remained motile for 2–3 h. Motility of B. boops and D. sargus spermatozoa was reversibly suppressed in the seminal plasma, and in electrolyte and non-electrolyte solutions of 100–200 mosmol kg^?1. The trigger for motility activation was hyperosmolality (700–1000 mosmol kg^?1). Motility of M. barbatus and T. mediterraneus sperm was only partly suppressed in the seminal plasma since freshly collected semen contained about 25–50% locally motile spermatozoa. When sperm was activated immediately after collection with electrolyte and non-electrolyte solutions of 700–1000 mosmol kg^?1 spermatozoa moved progressively. The motility of those spermatozoa which had not yet been motile after collection was completely and reversibly suppressed in M. barbatus at osmolalities of 1200 mosmol kg^?1, and at osmolalities of 100–200 mosmol kg^?1 in T. mediterraneus . Therefore two triggers were necessary for initiation of motility. The nature of the first trigger was uncertain, the second trigger was a switch to hypoosmolality in M. barbatus and to hyperosmolality in T. mediterraneus . The sperm organisation of B. boops, D. sargus, M. barbatus and T. mediterraneus revealed species-specific parameters which could not be related with the sperm motility behaviour.     

Urinary bladder, ionic composition of seminal fluid and urine with characterization of sperm motility in tench (Tinca tinca L.)

A small urinary bladder attached to the seminal duct in caudal part of the abdominal cavity was registered for the first time in dissected males of tench. The urinary bladder wall was of whitish color and the bladder contained 0.5–2 ml of urine. When collected in the experiment, the tench sperm was white‐colored. Spermatozoa density is highly variable due to contamination by urine, and the latter additionally activates spontaneous motility of the spermatozoa. Seminal fluid contains ions such as Na⁺ (18.4 ± 1.3 mm ), K⁺ (1.9 ± 0.6 mm ), Ca²⁺ (0.6 ± 0.2 mm ) and Mg²⁺ (0.5 ± 0.1 mm ), leading to osmolality of 230 ± 82 mOsmol kg^?1 depending on the dilution by urine. Urea was detected in urine samples uncontaminated by sperm with an osmolality of 85 ± 58 mOsmol kg^?1. Urine also contained high concentrations of ions such as Na⁺ (30.9 ± 8.9 mm ), K⁺ (4.3 ± 2.9 mm ), Ca²⁺ (0.9 ± 0.5 mm ) and Mg²⁺ (0.6 ± 0.2 mm ). The spontaneous sperm activation by urine was up to 100%, but could be prevented by collection in an immobilizing solution. Motility was observed for 90–100% spermatozoa just after their transfer to distilled water or in a swimming medium (SM, 30–45 mm KCl) with a velocity of 120–140 μm s^?1. A flagellar beat frequency of 60–70 Hz and forward motility lasted up to 80 s in distilled water, and up to 180 s in SM at room temperature.     

Spermatozoa motility and variation in the seminal plasma proteome of Eurasian perch (Perca fluviatilis) during the reproductive season

This study evaluated physiological and functional sperm parameters and the seminal plasma proteome of Eurasian perch (Perca fluviatilis) over the course of their reproductive season. Spermatozoa velocity (169.56 ± 6.53 to 158.5 ± 7.4 µm sec^?1), percent motility (95.89 ± 4.28% to 89.55 ± 4.5%), and osmolality of seminal plasma (290 ± 5 to 297 ± 12 mOsmol kg^?1) remained stable throughout the reproductive season. Milt volume and protein concentration of seminal plasma gradually increased and reached the highest values late in the reproductive period. Spermatozoa concentration peaked in the mid‐reproductive season (66.90 ± 13 × 10⁹ spermatozoa ml^?1) and decreased towards the end (54 ± 10 × 10⁹ spermatozoa ml^?1). A proteomic analysis of seminal plasma using two‐dimensional polyacrylamide gel electrophoresis revealed 10 protein spots significantly altered over the course of the reproductive season. Subsequent protein characterization suggested that time in the reproductive season predominantly affected proteins involved in membrane trafficking, organization, cell motility, and oxido‐reductase activity. This study provides new data on physiological properties of sperm and protein patterns of seminal plasma over the course of the reproductive season that should be considered in the development of methods for artificial reproduction of perch. Mol. Reprod. Dev. 79: 879–887, 2012. © 2012 Wiley Periodicals, Inc.     

The effects of environmental salinity on the growth and physiology of totoaba Totoaba macdonaldi and shortfin corvina Cynoscion parvipinnis

Totoaba Totoaba macdonaldi and shortfin corvina Cynoscion parvipinnis, were acclimated and reared together at salinities of 0, 2, 5, 10, 20 and 35 for 56 days. Initial overall mean ± s.d . body masses of 67·6 ± 7·1 g T. macdonaldi and 37·3 ± 3·1 g C. parvipinnis increased to final overall masses of 217·4 ± 30·3 and 96·5 ± 16·5 g, respectively, at the end of the study. Totoaba macdonaldi was not able to tolerate salinities of 0 and 2 and C. parvipinnis of 0. In contrast, both species had 100% survival at salinities ≥ 10. Somatic growth was highest not at natural seawater salinity of 35, but at 10. Plasma osmolality ranged from 172·5 to 417·0 mOsmol kg^?1 for T. macdonaldi and from 207·0 to 439·5 mOsmol kg^?1 for C. parvipinnis and varied in direct proportion to salinity. The estimated isosmotic salinities of T. macdonaldi and C. parvipinnis were 12·3 and 13·4, respectively. Cynoscion parvipinnis reared at two had significantly lower plasma lysozyme activity (95·0 Units ml^?1) than fish held at salinities from 5 to 35 (ranging from 215·0 to 355·0 Units ml^?1), but without clear trends over this range. Blood neutrophil oxidative radical production (NBT) (ranging from 3·9 to 6·7 mg ml^?1) had some significant differences among salinities, but these did not follow a clear pattern. For T. macdonaldi, neither lysozyme activity nor NBT was affected by salinity. Ash content of whole fishes varied directly and moisture content inversely, with salinity for both species.     

Effects of thymol and diphenyliodonium chloride against Campylobacter spp. during pure and mixed culture in vitro

Aims: To determine if the purported deaminase inhibitors diphenyliodonium chloride (DIC) and thymol reduce the growth and survivability of Campylobacter. Methods and Results: Growth rates of Campylobacter jejuni and Camp. coli were reduced compared to unsupplemented controls during culture in Muellar–Hinton broth supplemented with 0·25 μmol DIC or thymol ml^?1 but not with 0·01 μmol monensin ml^?1 or 1% ethanol. Recovery of Camp. jejuni and Camp. coli was reduced >5 log₁₀ CFU from controls after 24 h pure culture in Bolton broth supplemented with 0·25 or 1·0 μmol DIC ml^?1 or with 1·0 μmol thymol ml^?1. Similarly, each test Campylobacter strain was reduced >3 log₁₀ CFU from controls after 24 h mixed culture with porcine faecal microbes in Bolton broth supplemented with 0·25 or 1·0 μmol DIC ml^?1 or with 1·0 μmol thymol ml^?1. Treatments with 0·25 μmol thymol ml^?1, 0·01 μmol monensin ml^?1 or 1% ethanol were less effective. Ammonia production during culture or incubation of cell lysates was reduced by 0·25 or 1·0 μmol DIC ml^?1 but only intermittently reduced, if at all, by the other treatments. Conclusions: Diphenyliodonium chloride and thymol reduced growth, survivability and ammonia production of Camp. jejuni and Camp. coli. Significance and Impact of the Study: Results suggest a potential physiological characteristic that may be exploited to develop interventions.     

The effects of light intensity and algae-induced turbidity on feeding behaviour of larval striped trumpeter

Striped trumpeter larvae reared in algal cell‐induced turbid water (greenwater) fed equally well in clearwater in a light intensity range of 1–10 μmol s^‐1 m^‐2, when evaluated in terms of both the proportion of larvae feeding and larval feeding intensity. An ontogenetic improvement in photopic visual sensitivity of larvae was indicated by improved feeding at 0·1 μmol s^‐1 m^‐2, from 26±5% of larvae feeding and 0·027±0·005 rotifers consumed per feeding larva min^‐1 on day 8, to 96±2% and 0·221±0·007 rotifers consumed larva^‐1 min^‐1 on day 23 post‐hatching. Algal cell‐induced turbidity was shown to reduce incident irradiance with depth, indicated by increasing coefficients of attenuation (1·4–33·1) with increasing cell densities (0–2×10⁶ cells ml^‐1), though light intensities in the feeding experiment test chambers, at the algal cell densities tested, were within the optimal range for feeding (1–10 μmol s^‐1 m^‐2). Algae‐induced turbidity had different effects on larval feeding response dependent upon the previous visual environment of the larvae. Young larvae (day 9 post‐hatching) reared in clearwater showed decreased feeding capabilities with increasing turbidity, from 98±1% feeding and 0·153±0·022 rotifers consumed larva^‐1 min^‐1 in clearwater to 61±10% feeding and 0·042±0·004 rotifers consumed larva^‐1 min^‐1 at 56 NTU, while older clearwater reared larvae fed well at all turbidities tested. Likewise, greenwater reared larvae had increased feeding capabilities in the highest algal cell densities tested (32 and 66 NTU) compared with those in low algal cell density (6 NTU), and clearwater (0·7 NTU) to which they were naïve.     

COMPARISON OF DEVELOPMENT AND PHOTOSYNTHETIC GROWTH FOR FILAMENT CLUMPS AND REGENERATED MICROPLANTLET CULTURES OF AGARDHIELLA SUBULATA (RHODOPHYTA, GIGARTINALES)

Two axenic, in vitro liquid suspension cultures were established for Agardhiella subulata (C. Agardh) Kraft et Wynne, and their growth characteristics were compared. This study illustrated how reliable routes for the development of suspension cultures of macrophytic red algae of terete thallus morphology can be achieved for biotechnology applications. Undifferentiated filament clumps of 2–8 mm diameter were established by induction of callus-like tissue from thallus explants, and lightly branched microplantlets of 2–10 mm length were established by regeneration of filament clumps. The filament clumps were susceptible to regeneration. Adventitious shoot formation was reliably induced from 40% to 70% of the filament clumps by gentle mixing at 100 rev min^?1 on an orbital shaker. The specific growth rate of the microplantlets was higher than the filament clumps in nonagitated well plate culture (4%–6% per day for microplantlets vs. 2%–3% per day for filament clumps) at 24° C and 8–36 μmol photons·m^?2·s^?1 irradiance (10:14 h LD cycle) when grown on ASP12 artificial seawater medium at pH 8.6–8.9 with 20%–25% per day medium replacement. Oxygen evolution rate vs. irradiance measurements showed that relative to the filament clumps, microplantlets had a higher maximum specific oxygen evolution rate (P_o,max= 0.181 ± 0.035 vs. 0.130 ± 0.023 mmol O₂·g^?1 dry cell mass·h^?1), but comparable respiration rate (Q_o= 0.040 ± 0.013 vs. 0.033 ± 0.017 mmol O₂·g^?1 dry cell mass·h^?1), compensation point (I_c= 3.8 ± 2.4 vs. 5.7 ± 1.2 μmol photons·m^?2·s^?1), and light intensity at 63.2% of saturation (I_k= 17.5 ± 3.9 vs. 14.9 ± 2.6 μmol photons·m^?2·s^?1). The microplantlet culture was more suitable for suspension culture development than the filament clump culture because it was morphologically stable and exhibited higher growth rates.     

Maximum sustainable speed, energetics and swimming kinematics of a tropical carangid fish, the green jack Caranx caballus

Maximum sustained swimming speeds, swimming energetics and swimming kinematics were measured in the green jack Caranx caballus (Teleostei: Carangidae) using a 41 l temperature‐controlled, Brett‐type swimming‐tunnel respirometer. In individual C. caballus [mean ±s.d. of 22·1 ± 2·2 cm fork length (L_F), 190 ± 61 g, n = 11] at 27·2 ± 0·7° C, mean critical speed (U_crit) was 102·5 ± 13·7 cm s^?1 or 4·6 ± 0·9 L_F s^?1. The maximum speed that was maintained for a 30 min period while swimming steadily using the slow, oxidative locomotor muscle (U_max,c) was 99·4 ± 14·4 cm s^?1 or 4·5 ± 0·9 L_F s^?1. Oxygen consumption rate (M in mg O₂ min^?1) increased with swimming speed and with fish mass, but mass‐specific M (mg O₂ kg^?1 h^?1) as a function of relative speed (L_F s^?1) did not vary significantly with fish size. Mean standard metabolic rate (R_S) was 170 ± 38 mg O₂ kg^?1 h^?1, and the mean ratio of M at U_max,c to R_S, an estimate of factorial aerobic scope, was 3·6 ± 1·0. The optimal speed (U_opt), at which the gross cost of transport was a minimum of 2·14 J kg^?1 m^?1, was 3·8 L_F s^?1. In a subset of the fish studied (19·7–22·7 cm L_F, 106–164 g, n = 5), the swimming kinematic variables of tailbeat frequency, yaw and stride length all increased significantly with swimming speed but not fish size, whereas tailbeat amplitude varied significantly with speed, fish mass and L_F. The mean propulsive wavelength was 86·7 ± 5·6 %L_F or 73·7 ± 5·2 %L_T. Mean ±s.d . yaw and tailbeat amplitude values, calculated from lateral displacement of each intervertebral joint during a complete tailbeat cycle in three C. caballus (19·7, 21·6 and 22·7 cm L_F; 23·4, 25·3 and 26·4 cm L_T), were 4·6 ± 0·1 and 17·1 ± 2·2 %L_T, respectively. Overall, the sustained swimming performance, energetics, kinematics, lateral displacement and intervertebral bending angles measured in C. caballus were similar to those of other active ectothermic fishes that have been studied, and C. caballus was more similar to the chub mackerel Scomber japonicus than to the kawakawa tuna Euthynnus affinis.     

Lysozyme activities and immunoglobulin concentrations in seminal plasma and spermatozoa of different teleost species and indications on its significance for sperm function

The occurrence of lysozyme and immunoglobulin (Ig) in semen of different teleost species (brown trout—Salmo trutta, perch—Perca fluviatilis, burbot—Lota lota) was studied. In all investigated species lysozyme activities (1.13-1.45 U ml⁻¹) and Ig concentrations (T-Ig: 1.11-1.61 μg ml⁻¹, IgG [measured only in brown trout]: 1.49 μg ml⁻¹) were detected in seminal plasma. Ig was also found in spermatozoa (T-Ig: 0.234-0.357 μg/g protein, IgG: 0.198 μg ml⁻¹) while spermatozoal lysozyme activities were low and fluctuating (0.093-0.164 U/g protein). In Salmo trutta lysozyme activities and immunoglobulin levels were compared between semen samples with high and low sperm motility as motility is an indicator for sperm fertility. Lysozyme activities were higher in seminal plasma of samples with high motility than in those with low motility while seminal plasma and spermatozoal immunoglobulin concentrations (T-Ig, IgG) were increased in samples with low motility in comparison to samples with high motility. Seminal plasma and spermatozoal IgG concentrations and seminal plasma lysozyme activities showed significant correlations with the sperm motility rate and swimming velocity. Moreover, lysozyme improved the viability of spermatozoa in in vitro experiments. Possible physiological meanings of these results are discussed.     

Effect of light intensity and eye development on prey capture by larval striped bass Morone saxatilis

The efficacy of visual and non-visual feeding among pelagic striped bass Morone saxatilis larvae adapted to a turbid estuary was determined in the laboratory in clear water. Capture of Artemia salina (density 100 l⁻¹) was significantly affected by the interaction between age of larvae (range: 8–25 days post-hatch, dph) and light intensity (range: 0–10·6 μmol s⁻¹ m⁻² at the water surface). Visual feeding by larvae aged 9–11 dph was highest in dim light (0·086–0·79 μmol s⁻¹ m⁻²), with fish capturing up to 5 prey larva⁻¹ h⁻¹. As the larvae grew, prey capture in brighter light improved, associated with an increasing proportion of twin cone photoreceptors and improving ability of the retina to light- and dark-adapt. By age >22 dph, mean prey capture was greatest at highest light intensities (0·79 and 10·6 μmol s⁻¹ m⁻²) exceeding 100 prey larva⁻¹ h⁻¹. Incidence of feeding larvae generally improved as the larvae grew, reaching >80% in all light intensities from 16 dph onwards. The lower threshold for visual feeding, between 0·0084 and 0·03 μmol s⁻¹ m⁻², remained constant as the larvae grew, despite an increasing density of rod photoreceptors. Below this threshold, non-visual feeding was evident at a low rate (<6 prey larva⁻¹ h⁻¹) that was independent of larval age.     

Relationships between reproductive characteristics in male Vimba vimba L. and the effects of osmolality on sperm motility

In Vimba vimba, GSI, sperm volume, and spermatozoa concentration range from 3.4-7.4 %, 0.1-1.1 ml, and 13.3-34.8 × 10⁹ spz ml⁻¹, respectively. Gonad mass (r = 0.90) and sperm volume (r = 0.35) significantly correlated with weight of males. Significant correlation was also found between gonad mass and length of males (r = 0.85). Sperm motility (r = 0.99) and velocity (r = 098) significantly decreased after activation in Tris-HCl 20 mM, pH 8.5. Osmolality of the seminal plasma was 273.2 mOsmol kg⁻¹. Sperm motility and velocity were significantly affected by the osmolality of the activation medium (P < 0.01). Hyper-osmolality compared to seminal plasma osmolality totally suppressed the sperm activation. At 15 s post-activation, the sperm motility significantly decreased at 240 mOsmol kg⁻¹ in KCl or NaCl media. The highest sperm motility and velocity (at 60 s post-activation) were observed at 200 mOsmol kg⁻¹ in NaCl, KCl, or sucrose media. In all treatments, the tip of the flagellum of spermatozoa became curled into a loop shape after activation of sperm in distilled water containing 20 mM Tris-HCl, pH 8.5 that shortened the flagellum.     

The influence of the length of time after hormonal treatment with [(d‐Ala6, Pro9 NEt)‐mGnRH+metoclopramide] i.e. Ovopel on barbel Barbus barbus (L.) milt quality and quantity indicators

The study purpose was the analysis of barbel Barbus barbus (L.) milt quality and quantity with regard to the time following stimulation with [(D‐Ala⁶, Pro⁹NEt)‐mGnRH+metoclopramide] i.e. Ovopel. Selected parameters such as total volume of milt (TVM, ml), volume of milt per kg of body weight (VOM, ml kg^?1 b.w.), sperm concentration (×10⁹ ml^?1), total sperm production (TSP, ×10⁹), osmolality (mOsm kg^?1) and pH of seminal plasma were determined. Sperm motility was analyzed by the CASA system, i.e. the percentage of sperm motility (MOT, %) and their progressive motility (PRG, %), curvilinear velocity (VCL, μm s^?1) and straight line velocity (VSL, μm s^?1), amplitude of lateral head displacement (ALH, μm), and beat cross frequency (BCF, Hz). Milt was collected from 12 specimens (N = 12), with the first portion obtained 12 h following treatment with Ovopel (1 granule kg^?1 b. w.). Subsequent portions of milt were taken at 24 h intervals, i.e. after 36, 60, 84, 108, and 132 h. The control group (Control, N = 12) was injected with 0.9% NaCl at 0.5 ml kg^?1b.w. from which milt was taken 12 h post injection. The highest TVM, VOM and TSP values were recorded 12 h after Ovopel treatment (3.2 ± 0.7 ml, 36.7 ± 10.5 ml kg^?1 b.w. and 39.1 ± 9.4 × 10⁹, respectively); lowest values were recorded after 132 h (0.8 ± 0.4 ml, 11.1 ± 6.5 ml kg^?1b.w. and 13.7 ± 7.5 × 10⁹, respectively). The highest seminal plasma osmolality values (300.0 ± 42.6 mOsm kg^?1) as well as the lowest sperm concentration (12.5 ± 1.5 × 10⁹ ml^?1) were observed 12 h after Ovopel treatment. No significant differences in the percentage of sperm motility (MOT) were noted during any of the periods after hormonal stimulation, however, a change in the character of their movement (PRG) was observed. The lack of significant differences (P > 0.05) in VCL and VSL values between 12 h and 60–132 h indicates that the lengthening of time does not lead to a decrease in sperm velocity and, therefore, is not likely to have a negative impact on their quality. The highest ALH (1.9 ± 0.2 μm) and BCF (11.5 ± 1.1 Hz) values were observed when the effect of stimulation was most noticeable, i.e. 12 h after Ovopel treatment. Based on the total milt volume and sperm production, the best time for milt collection from barbel is 12 h post‐hormonal treatment; 84 h post‐hormonal treatment, TVM, VOM, TSP and some CASA parameters decreased, which suggest the same aging process in sperm.     

Seasonal nitrate physiology of Macrocystis integrifolia Bory

Ambient sea-water nitrate and tissue nitrogen (ethanol soluble nitrate and amino acids, as well as total nitrogen) of Macrocystis integrifolia Bory were monitored over a 2-yr period in Bamfield, Vancouver Island, British Columbia. Sea-water nitrate varied from a high of 12 μmol · 1^?1 (individual values as high as 23 μmol · 1^?1 were recorded) in late winter to below detection limits for most of the summer. Tissue nitrate and total nitrogen paralleled the ambient nitrate levels and showed summer minima and winter maxima (from 0 to 70 μmol · g fresh wt^?1 for nitrate and from 0.8 to 2.9% of dry wt for total N). The nitrate uptake capacity was inversely proportional to tissue nitrate concentration and, furthermore, was much higher for subapical surface blades (60–70 nmol · cm^?2 · h^?1) than for older, deeper blades (5–10 nmol · cm^?2 · h^?1). Nitrate uptake by subapical blade disks in summer is apparently higher in dark (1.0–1.7 μmol · g fresh wt^?1 · h^?1) than in light (0.6–1.3 μmol · g fresh wt^?1 · h^?1) and the data obtained in 36–108 h experiments indicate nitrate pool sizes of 30–90 μmol · g fresh wt^?1. These pools are $\text{2}\text{3}$ to nearly full in winter. Ammonium does not inhibit nitrate uptake. It is taken up and apparently utilized much faster than nitrate and it may well be an important source of nitrogen for marine macrophytes.     

Respiratory-cardiovascular physiology and chloroethane gill flux in the channel catfish, Ictalurus punctatus

A fish respirometer-metabolism chamber was used to obtain in vivo respiratory-cardiovascular and chloroethane gill flux data on transected channel catfish (Ictalurus punctatus). Methods used for spinal transection, attachment of an oral membrane (respiratory mast), placement and attachment of blood cannulas and urine catheters are described. Respiratory physiology, cardiac output and chemical extraction efficiencies for 1,1,2,2-tetrachloroethane (TCE), pentachloroethane (PCE), and hexachloroethane (HCE) were determined on 419–990 g catfish. The overall mean values (± s.d.) for ventilation volume (Q_v), effective respiratory volume (Q_w), oxygen consumption (Vo₂ and percentage utilization of oxygen (U) were 17-3 ±4–71 h^?1 kg^?1, 9·8±l·71 h^?1 kg^?1, 71·6±12·5mg h^?1 kg^?1, and 49± 10%, respectively, while cardiac output calculated via the Fick Method was 2·4±0·61 h^?1 kg^?1. Additional measurements were made on ventilation rate (V_r), total plasma protein, haematocrit (Hct), and urine volume; while both arterial and venous blood were analysed for pH, oxygen partial pressure (P02), carbon dioxide partial pressure (Pco₂), total oxygen (To₂), total carbon dioxide (Tco₂) and total ammonia (TAMM). Physiological measurements taken at 24 h were not significantly different from those taken at 48 h and indicated no deterioration of the in vivo preparation. All of these values agreed well with literature values on UTitransected channel catfish, except for Hct which was lower for cannulated animals used in this study. Overall, these data provide strong support for the use of transected channel catfish for in vivo collection of physiological and chemical gill flux data. The mean initial chemical extraction efficiencies for TCE, PCE and HCE were 41, 61 and 73%, respectively. Chemical clearances (Cl_X) for these same three chemicals were 5·9, 9·3 and 10·8 1 h^?1 kg^?1, respectively. The approximate 1: 1 relationship between effective respiratory volume (Q_w) and chemical clearance (Cl_x) indicated that branchial uptake of PCE and HCE was water flow-limited. Chemical gill flux observed for channel catfish and chloroethanes was similar to that observed for rainbow trout in previous studies and provided further support for the flow-limited model of chemical flux across fish gills.     

Ecophysiology of the hypotonic response in the salt-tolerant charophyte alga Lamprothamnium papulosum

The ecophysiology of the hypotonic response was studied in the charophyte alga, Lamprothamnium papulosum, which was grown in a marine (SW; 1072 mosmol kg^–1) and a brackish (1/2 SW; 536 mosmol kg^–1) environment. The cells produced an extracellular mucilage identified by histochemical staining as a mixture of sulphated and carboxylated polysaccharides. The thickness and chemical composition of the mucilage layer was a function of environmental salinity and cell age. Mucilage progressively increased in thickness from the apex (9 SW cells: 12·6 ± 1·8 μm; 15 1/2 SW cells: 4·8 ± 0·7 μm) to the base of the plants (15 SW cells: 44·8 ± 3·3 μm; nine 1/2 SW cells: 23·8 ± 2·5 μm); with a corresponding increase in the sulphated proportion. The mucilage was significantly thicker in SW plants. Hydraulic conductivity (Lp) at the apex of SW plants, measured by transcellular osmosis, was 8·3 × 10^–13 m s^–1 Pa^–1. This was close to Lp of freshwater Chara (8·5 × 10^–13 m s^–1 Pa^–1) which lacked mucilage. Basal SW cells with thicker mucilage had a smaller apparent Lp of 3·5 × 10^–13 m s^–1 Pa^–1. The electrophysiology of the resting state and hypotonic response was compared in cells from the two environments based on current/voltage (I/V) analysis. The resting potential difference (PD) and conductance differed (11 SW cells: – 102·4 ± 10·1 mV, eight SW cells: 18·6 ± 2·4 S m^–2; 19 1/2 SW cells: –125·7 ± 5·9 mV, 8·3 ± 0·8 S m^–2). The type of cellular response to a hypotonic shock (decrease of 268 mosmol kg^–1) also differed. In 1/2 SW plants, only the apical cells with thin mucilage responded classically with depolarization, conductance increase, Ca²⁺ influx, cessation of cytoplasmic streaming, and K⁺ and Cl^– effluxes. Older cells making up the bulk of the plants responded with depolarization, but continued cytoplasmic streaming, and had only a small increase in conductance; or depolarized transiently without altering the I/V profile, conductance or streaming speed. Most cells remained depolarized and in the K⁺ state 1 h post-shock. Cells treated with the K⁺ channel blocker tetraethylammonium chloride also depolarized and remained depolarized. The SW cells depolarized but otherwise responded minimally to a 268 mosmol kg^–1 drop in osmolarity and required a further 268 mosmol kg^–1 down-step to elicit a change in the conductance. A spectrum of responses was measured in successively older and more mucilaginous cells from the same marine plant. We discuss the ecophysiological significance of the mucilage layer which modulates the cellular response to osmotic shock and which can be secreted to different degrees by plants inhabiting environments of different salinity.     

Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss,Walbaum)

Aims: To develop probiotics for the control of vibriosis caused by Vibrio anguillarum and Vibrio ordalii in finfish. Methods and Results: Kocuria SM1, isolated from the digestive tract of rainbow trout, was administered orally to rainbow trout (Oncorhynchus mykiss) for 2 weeks at a dose equivalent to c. 10⁸ cells per g of feed and then challenged intraperitoneally with V. anguillarum and V. ordalii. Use of SM1 led to a reduction in mortalities to 15–20% compared to 74–80% mortalities in the controls. SM1 stimulated both cellular and humoral immune responses in rainbow trout, by elevation of leucocytes (5·5 ± 0·8 × 10⁶ ml^?1 from 3·7 ± 0·8 × 10⁶ ml^?1), erythrocytes (1·2 ± 0·1 × 10⁸ ml^?1 from 0·8 ± 0·1 × 10⁸ ml^?1), protein (23 ± 4·4 mg ml^?1 from 16 ± 1·3 mg ml^?1), globulin (15·7 ± 0·2 mg ml^?1 from 9·9 ± 0·1 mg ml^?1) and albumin (7·3 ± 0·2 mg ml^?1 from 6·1 ± 0·1 mg ml^?1) levels, upregulation of respiratory burst (0·05 ± 0·01 from 0·02 ± 0·01), complement (56 ± 7·2 units ml^?1 from 40 ± 8·0 units ml^?1), lysozyme (920 ± 128·8 units ml^?1 from 760 ± 115·3 units ml^?1) and bacterial killing activities. Conclusions: Kocuria SM1 successfully controlled vibriosis in rainbow trout, and the mode of action reflected stimulation of the host innate immune system. Significance and Impact of the Study: Probiotics can contribute a significant role in fish disease control strategies, and their use may replace some of the inhibitory chemicals currently used in fish farms.     

Basic physico-chemical parameters of milt from sea trout (Salmo trutta m. trutta), brook trout (Salvelinus fontinalis) and rainbow trout (Oncorhynchus mykiss)

The aim of the study was to compare the physico‐chemical parameters of milt from sea trout (Salmo trutta m. trutta), brook trout (Salvelinus fontinalis) and rainbow trout (Oncorhynchus mykiss). Milt was collected by stripping and spermatozoa concentrations, were determined and compared with sperm motility and spermatocrit values along with seminal plasma indices (pH, osmolality, sodium, potassium, chlorine, calcium, magnesium, glucose and protein concentrations). The highest spermatozoa concentration of 22.3 ± 6.7 × 10⁹ ml^?1 was found in the sea trout milt, and was significantly different of those observed in brook trout (11.9 ± 3.3 × 10⁹ ml^?1) and rainbow trout (10.7 ± 4.4 × 10⁹ ml^?1). The values for pH and K⁺ did not differ significantly among species. The mean pH was 8.0 in the milt of each species and the K⁺ concentrations ranged from 24.8 ± 7.2 to 30.5 ± 7.6 mm L^?1. Considerable differences were determined for the Ca²⁺ ions concentrations. The highest value was found in sea trout (1.7 ± 0.3 mm L^?1), while in the rainbow trout it was 0.7 ± 0.5 and in the brook trout 0.4 ± 0.1 mm L^?1. The most pronounced differences were found in the glucose concentration cause of its unnaturally low concentration in rainbow trout of the mean value of 6.0 ± 15.2 mg L^?1. The mean value in sea trout and brook trout was 185.0 ± 172.4 and 231.2 ± 148.4 mg L^?1 respectively. For all species, protein mean values were below 1.3 g L^?1. The mean osmolality was between 230.6 ± 98.6 and 272.0 ± 26.4 mOsm kg^?1 in the species studied. No correlation was found between any components determined in milt and the spermatozoa motility (P > 0.05). The sperm concentration was positively correlated with the protein content in the milt of the three species studied, other less exhibited correlation was found.     

Aerobic scope for activity in age 0 year Atlantic cod Gadus morhua

Key components of swimming metabolism: standard metabolism (R_s), active metabolism (R_a) and absolute aerobic scope for activity (R_a–R_s) were determined for small age 0 year Atlantic cod Gadus morhua. Gadus morhua juveniles grew from 0·50 to 2·89 g wet body mass (M_WB) over the experimental period of 100 days, and growth rates (G) ranged from 1·4 to 2·9% day^?1, which decreased with increasing size. Metabolic rates were recorded by measuring changes in oxygen consumption over time at different activity levels using modified Brett‐type respirometers designed to accommodate the small size and short swimming endurance of small fishes. Power performance relationships were established between oxygen consumption and swimming speed measurements were repeated for individual fish as each fish grew. Mass‐specific standard metabolic rates () were calculated from the power performance relationships by extrapolating to zero swimming speed and decreased from 7·00 to 5·77 μmol O₂ g^?1 h^?1, mass‐specific active metabolic rates () were calculated from extrapolation to maximum swimming speed (U_max) and decreased from 26·18 to 14·35 μmol O₂ g^?1 h^?1 and mass‐specific absolute scope for activity was calculated as the difference between active and standard metabolism () and decreased from 26·18 to 14·35 μmol O₂ g^?1 h^?1 as M_WB increased. Small fish with low R_s had bigger aerobic scopes but, as expected, R_s was higher in smaller fish than larger fish. The measurements and results from this study are unique as R_s, R_a and absolute aerobic scopes have not been previously determined for small age 0 year G. morhua.