Short-term thermal acclimation induces adaptive changes in the inner mitochondrial membranes of fish skeletal muscle

In the oxidative muscles (musculi laterales superficiales) of crucian carp Carassius carassius acclimated for 6 weeks to either 5 or 25° C, the volume density and the surface density of fibres per tissue did not differ significantly between the control and experimental groups. The correlation ratio (μ²) for these values was below 50, 39·3 and 43·9 respectively. After acclimation to 5° C, the surface density of outer mitochondrial membrane per fibre increased significantly from 0·93 to 1·23m² cm⁻³ in the summer population but dropped from 0·94 to 0·67 m² cm⁻³ in the winter population. The surface density of outer mitochondrial membrane per mitochondrion increased from 3·24 to 4·52 m² cm⁻³ in summer fish. After acclimation to 25° C, the surface density of inner mitochondrial membranes per muscle fibre decreased from 4·04 to 1·79 m² cm⁻³ in summer fish and from 3·86 to 1·07 m² cm⁻³ in winter fish. The surface density of inner mitochondrial membranes per mitochondrion increased from 14·17 to 15·60 m²cm⁻³ in summer fish but dropped from 13·91 to 10·67 m² cm⁻³ in winter fish. Correlation matrices demonstrate a negative correlation of the surface density of outer mitochondrial membrane per mitochondrion with the volume density of mitochondria per fibre and temperature, suggesting cold-induced proliferation of small mitochondria. It was concluded that short-term cold acclimation increased surface area of the inner mitochondrial membranes in summer fish.     

Lipid content and fatty acid composition of target tissues in wild Perca fluviatilis females in relation to hepatic status and gonad maturation

Variation in liver ultra‐structure and composition in relation to energy mobilization was investigated in female perch Perca fluviatilis from the Meuse River between August 2001 and June 2002. In April, just before spawning, the lipo‐somatic index ( I _F) was 0·3%, the gonado‐somatic index ( I _G) was 28% and the total lipid content of the liver was 2·53%. The average areas of lipid droplets and mitochondria were 0·05 and 0·06 μm², respectively. Glycogen supply reached 7·9% of the total area of the hepatocyte. During the sexual resting period, females accumulated energy in perivisceral fat and in the liver to reach 1·6% I _F and 4·85% of liver lipid content in August with lipid droplets average size of 0·09 μm² and glycogen average area of 15%. Liver cells contained a weakly developed rough endoplasmic reticulum (RER) and a great number of small mitochondria (average size 0·02 μm²). The I _G was 0·6% at this time. During the whole annual cycle, the average lipid content of female liver never exceeded 3·9 ± 1·9%. The concentration of docosahexaenoic (DHA), linolenic and linoleic acids increased in mature gonads while linolenic and linoleic acids decreased in the liver during the same period. Fatty acid composition of muscles of perch was characterized by a high content of DHA.     

Effects of cadmium chloride on the development of rainbow trout Oncorhynchus mykiss early life stages

The sub-chronic (28–56 days) effects of exposure to low concentrations of cadmium (Cd; 0·05, 0·25, 0·50 and 2·50 μg l⁻¹) shortly following fertilization on embryos, larvae and juvenile rainbow trout Oncorhynchus mykiss were examined. Premature hatching occurred at lower concentrations (0·05 and 0·25 μg l⁻¹ Cd), however, delayed hatching was seen in the 2·50 μg l⁻¹ Cd group, with >90% of hatching occurring on the last day of the hatching period. Larval growth was negatively affected by Cd exposure in a concentration-dependent manner. Larvae exposed to 2·50 μg l⁻¹ Cd were 13·9 ± 0·8% shorter in total length ( L _T) and weighed 22·4 ± 3·5% (mean ± s . e .) less than controls at the end of the exposure period. Plasma sex steroid concentrations (oestradiol in juvenile females and 11-ketotestosterone in juvenile males) were elevated (four- to 10-fold over controls) in exposed fish in both males and females, following 28 days of exposure to 0·05, 0·25 and 0·50 μg l⁻¹ Cd, respectively. These results suggest that environmentally realistic concentrations (in the μg l⁻¹ range) of Cd can affect the development of O. mykiss impacting embryos, larvae and juvenile fish.     

Seasonal patterns of nitrogen fixation in termites

1. Termite nitrogenase activity was highest in autumn and spring (≈ 3 μg N₂ fixed termite fresh mass (g)^–1 day^–1) and lowest in winter and summer (≈ 0·8 μg N₂ fixed termite fresh mass (g)^–1 day^–1).
2. The nitrogenase activity of worker termites was significantly higher than all other castes (1·58 ± 0·27 μg N₂ fixed termite fresh mass (g)^–1 day^–1).
3. Worker termites constituted the largest proportion of all the castes throughout the study period (≈ 90%).
4. The localized input of fixed nitrogen by termites may reach 15·3 mg N log^–1 day^–1 and 5·6 g N log^–1 year^–1.     

Osmoregulation during the development of glass eels and elvers

Drinking rates in glass eels and elvers of the European eel increased with environmental acclimation salinity from 0·07±0·02 (FW) to 0·70±0·09 μl g-¹ h-¹ (SW) at month 1 and from 1·12±0·42 (FW) to 12·85±1·05 ± l g-¹ h-¹ (SW) at month 5. Drinking rates increased with time in both FW and SW groups. FW acclimated eels when challenged acutely with SW increased drinking rate rapidly immediately upon transfer (0–15 min) and the magnitude of this response increased with developmental time from month 1 to month 5.     

Relationships between biovolume and carbon and nitrogen content of bacterioplankton

Abstract Cell volume, carbon and nitrogen content were determined for bacteria grown in batch cultures in water samples collected at five localities in western Florida, USA. Cultures were set up by inoculating 0.2 μm filtered water with 2.5 to 7.0% of 1.0 μm filtered water. Biovolumes of the bacteria were measured by epifluorescence photomicrography. Bacterial carbon and nitrogen contents were determined with a CHN analyser. During incubations, bacterial volumes doubled from 0.070±0.037 μ m³(mean ± S.E.) to 0.153 ± 0.036 μ m³ at early stationary phase. Bacterial C:N ratios ranged between 2.8 and 10.3, with a mean of 6.5, and were inversely correlated with cell volumes. Conversion factors for volume to carbon and nitrogen content were relatively high and variable, ranging from 0.21 to 161 pg C μm⁻³ (mean: 0.72 pg C μm⁻³) and from 0.05 to 0.25 pg N μm⁻³ (mean: 0.12 pg N μm⁻³). Small cells contained more C and N per unit volume than did large cells. The data suggested that biovolume to biomass conversion factors may be higher than previously thought and may be highly variable both temporally and geographically.     

Monoamine oxidase activity in kidney and heart of Piaractus mesopotamicus (Holmberg)

The values of Michaelis–Menten constant (K_M) and maximum velocity (V_MAX) for kidney and heart monoamine oxidase (MAO) from pacu Piaractus mesopotamicus were determined. The mean ± s . e . K_M values were 17·28 ± 2·27 μM for kidney and 15·38 ± 1·86 μM for heart. MAO activities were 111·60 ± 3·25 and 15·12 ± 0·30 nmols min⁻¹ g⁻¹ of wet tissue for kidney and heart, respectively. In addition, MAO inhibitory studies in these two tissues indicate that this enzyme may be a different isoform of MAO.     

A method for measuring oxygen consumption in isolated perfused gills

A method is described for measuring respiration in isolated perfused flounder gills experiencing pressures and flows similar to those seen in vivo . Mean oxygen consumption of 13 preparations bathed and perfused in identical saline was 5·00 ± 0·75 (s.e.) μ mol h⁻¹ g wet⁻¹, whilst that of five preparations perfused with saline but bathed in sea water (32 mg l⁻¹) was 12·06±2·39 (s.e.) μmol h⁻¹ g wet⁻¹. The oxygen consumption of the seawater bathed gills was significantly higher (P<0·05) than that in saline bathed gills. These results provide direct evidence both of the high metabolic activity of the gill under normal perfusion conditions and of the increased energy expenditure of the giil in hyperosmotic, compared to isosmotic, environments.     

Use of a Coulter Counter to Detect Discrete Changes in Cell Numbers and Volume During Growth of Escherichia coli

Escherichia coli could be detected at a concentration of 10⁵ cells/ml. Growth curves of the organism grown in filtered tryptone soya broth were made by taking viable and Coulter counts. Both curves were similar in shape and varied little during 10 h growth. Cell volume and volume distribution were seen to vary considerably however, volume reaching a stable minimum value of 0·61 μm³ after 8 h. The Coulter Counter was considered to be potentially useful for detecting significant bacteriuria.     

Effects of osmolality and ions on the motility of stripped and testicular sperm of freshwater-and seawater-acclimated tilapia, Oreochromis mossambicus

A significantly higher concentration of testicular spermatozoa was obtained from freshwater Oreochromis mossambicus (9·9×10⁹ spermatozoa ml⁻¹) than seawater O. mossambicus (4·6×10⁹ spermatozoa ml⁻¹). The mean osmolality of the urine of freshwater fish (78·5 mOsmol kg⁻¹) was significantly different from that of seawater fish (304·8 mOsmol kg⁻¹). The mean length of the mid-piece of the spermatozoa together with the tail was more variable in freshwater O. mossambicus (8·80±0·23μm) than in seawater specimens (8·27±0·18 μm). Stripped sperm of freshwater O. mossambicus was highly contaminated by urine which was a good activator of sperm motility in O. mossambicus held in both fresh and sea water. The osmolality for initiation of motility in freshwater O. mossambicus spermatozoa was from 0 to 333 mOsmol kg⁻¹ while for seawater O. mossambicus spermatozoa it was from 0 to 1022 mOsmol kg⁻¹. The optimum osmolality for motility was from 70 to 333 mOsmol kg⁻¹ for freshwater O. mossambicus spermatozoa and from 333 to 645 mOsmol kg⁻¹ for seawater fish. In freshwater O. mossambicus spermatozoa, the presence of 20 mM CaCl₂ increased the permissive osmolality of NaCl from 184 to 645 mOsmol kg⁻¹. For seawater O. mossambicus spermatozoa, solutions of NaCl devoid of CaCl₂ were unable initiate motility, but the addition of 1·5 to 30 mM CaCl₂ to the NaCl solution (0–934 mOsmol kg₁) had a full motility initiating effect.     

Physiological impact of sea lice on swimming performance of Atlantic salmon

Atlantic salmon Salmo salar were infected with two levels of sea lice Lepeophtheirus salmonis (0·13 ± 0·02 and 0·02 ± 0·00 sea lice g⁻¹). Once sea lice became adults, the ventral aorta of each fish was fitted with a Doppler cuff to measure cardiac output ( ̇ ), heart rate ( f _H) and stroke volume ( V _S) during swimming. Critical swimming speeds ( U _crit) of fish with higher sea lice numbers [2·1 ± 0·1 BL (body lengths) s⁻¹] were significantly lower ( P  < 0·05) than fish with lower numbers (2·4 ± 0·1 BL s⁻¹) and controls (sham infected, 2·6 ± 0·1 BL s⁻¹). After swimming, chloride levels in fish with higher sea lice numbers (184·4 ± 11·3 mmol l⁻¹) increased significantly (54%) from levels at rest and were significantly higher than fish with fewer lice (142·0 ± 3·7 mmol l⁻¹) or control fish (159·5 ± 3·5 mmol l⁻¹). The f _H of fish with more lice was 9% slower than the other two groups at U _crit. This decrease resulted in ̇ not increasing from resting levels. Sublethal infection by sea lice compromised the overall fitness of Atlantic salmon. The level of sea lice infection used in the present study was lower than has previously been reported to be detrimental to wild Atlantic salmon.     

Freeze-Fracture Study of the Trophozoite and the Cyst of Entamoeba histolytica

ABSTRACT The fine structure of the trophozoite and cyst of Entamoeba histolytica from the stool of a patient was compared using the freeze-fracture method. The intramembranous particles (IMP's) were heterogeneously distributed on the plasma membrane of the trophozoite and their density was 1139 ± 105/μm² on the P face and 27 ± 9/μm² on the E face. Particle-rich depressions and linear particle arrays, reported by other investigators on cultured trophozoites, were also observed on the P face while on the E face such special particle arrangement was not recognized. Particle-free, small protrusions were frequently observed on the P face of the trophozoite membrane. The existence of these protrusions is a new finding. In the cyst, the IMP's were also distributed heterogeneously on both the P and E faces of the plasma membrane. The density of the IMP's, however, was much lower than in the trophozoite: 6 ± 2/μm² on the P face and averaging less than 1/μm² on the E face. In freeze-fracture images, the plasma membrane of the cyst showed a variety of configurations from smooth to uneven or ridged surfaces. These morphological alterations of the plasma membrane may be attributed to the aging of the cyst. The thick wall of the cyst had a filamentous tri- or tetra-lamellar structure. The cytoplasm of the cyst was similar in structure to that of the trophozoite and the diameter of the nuclear pores was equal in both trophozoites and cysts.     

Difference in blood and water diffusion distance in gill lamellae of diploid and triploid tench Tinca tinca (L.)

Image analysis of sagittal sections of gill lamellae of diploid and triploid tench Tinca tinca revealed the blood and water diffusion distance in diploids (2·07 μm) to be significantly higher than that of their triploid siblings (1·46 μm; P < 0·01). Lamellae of diploids compared to triploids were found to be significantly shorter (105·84 v. 132·11 μm) and thicker (18·47 v. 14·21 μm; all at P < 0·05) than those of their triploid siblings but with similar mean sectional areas (1965·44 v. 1910·86 μm²).     

Territorial defence by the Brazilian damsel Stegastes fuscus (Teleostei: Pomacentridae)

Territorial defence by Brazilian damselfish Stegastes fuscus was studied in north-east Brazil through underwater observations. The fish mostly fed on Gellidium sp. and strongly defended a territory with a mean size of 2 m² (range 1·27–3·07 m²); its mean ± s.d. feeding rate was 8·73 ± 2·44 bites min⁻¹. Stegastes fuscus responded similarly to all intruding fish species, with aggressive behaviour becoming more pronounced only with an increase in the number of fishes in the sampled area. Analysis of attacks elicited by different fish species placed within S. fuscus ' territory, however, showed that it launched more attacks on conspecifics than on other fishes.     

Some morphometric and biochemical features of ready-to-migrate silver and pre-migratory yellow stages of the shortfin eel of south-eastern Australian waters

The mean total length ( L _T), mass and age of ready to migrate female silver shortfin eels Anguilla australis from the Hopkins River estuary and the mouth of the Merri River in south-eastern Australia, were 83·2 ± 1·2 cm, 1051 ± 51 g, and 17·2 ± 1·79 years, respectively. The eye index ( I _E) of the silver shortfin eels was < 5·2 (mean 7·64 ± 0·29) and differed significantly from that of the yellow shortfin eels collected from two other sites. The I _E increased with L _T (mm) and was related by log I _E= 2·656 log L _T6·925. The per cent moisture, protein and ash content of the liver of silver shortfin eels was significantly lower than in yellow shortfin eels, but lipid content was significantly higher in the former (35·5 ± 2·0%). The mean mass μg mg lipid ⁻) of saturates (230·4 ± 2·6 v . 181·7 ±2·6), monoenes (367·4 ± 6·3 v . 290·8 ± 8·9) and PUFA (177·3 ± 5·3 v . 159·7 ± 4·6) in muscle was significantly higher, and the great majority of individual fatty acids was found also in higher quantities in silver shortfin eels. In the liver, the PUFA found in the highest quantity was 22:6n-3, except in shortfin eels from Hopkins River estuary, and the amount of 18:2n-6 in the liver of silver shortfin eels was significantly higher than that in yellow shortfin eels but the reverse was true of 20:4n-6. In both muscle and liver tissues the saturate 16:0 and the monoene 18:ln-9 collectively accounted for >50% of all the fatty acids in the lipid.     

Quantitative Ultrastructural Investigations on Synaptogenesis in the Cerebellum and the Optic Tectum of Light-Reared and Dark-Reared Rainbow Trout (Salmo gairdneri Rich.)

The morphogenetic differentiation of synapses in the cerebellum and the optic tectum of darkand light- reared rainbow trout was investigated at critical stages of development. During normal differentiation the cerebellum is characterized by the appearance of 'indented', spinelike synapses. This type of synapses increases with age and prevails from day 60 on. At the same time the number of 'flat' synapses decreases. In the cerebellum the highest synaptic density (123 ± 12 synapses/1,000 μm²) is reached 30 days after hatching when the larvae begin to swim. The optic tectum is characterized by a preponderance of flat synapses in early postnatal and adult life; maximal synaptic density (66 ± 5 synapses/1,000 μm²) is reached 60 days after hatching when the larvae have reached optimal visual acuity.
Light deprivation causes a considerable and significant reduction in the number of synapses per unit area in the cerebellum and the optic tectum. The length of synaptic contacts do not change. If light-deprived, the density of synaptic vesicles decreases significantly in the optic tectum of a 25-day-old trout (74 ± 3 instead of 132 ± 7 vesicles/μm²). In the cerebellum this effect is absent.     

Generation of a reproducible nutrient-depleted biofilm of Escherichia coli and Burkholderia cepacia

An in vitro method of growing bacteria as a defined nutrient-depleted biofilm is proposed. The medium was defined nutritionally in terms of the quantitative composition and by the total amount of nutrient required to achieve a defined population size. Escherichia coli and Burkholderia cepacia were incubated on a filter support placed on a defined volume of solid medium. The change of biomass of the biofilm population was compared with the change in a planktonic culture. The size of the population in stationary phase was proportional to the concentration of limiting substrate up to 40 μmol cm⁻² glucose for E. coli and up to 2·7 × 10⁻⁹ mol cm⁻² iron for B. cepacia . Escherichia coli growing exponentially had a growth rate of μ = 0·30 h⁻¹ in a biofilm and μ = 0·96 h⁻¹ in planktonic culture. The growth rate, μ, for exponentially growing B. cepacia in a biofilm was 1·12 h⁻¹ and in planktonic culture 0·78 h⁻¹. This method allows the limitation of the size of a biofilm population to a chosen value.     

Symbiotic N2 fixation in a high Alpine grassland: effects of four growing seasons of elevated CO2

1. Increasing carbon dioxide concentration (E: 680 μl CO₂ litre^–1 vs ambient, A: 355 μl CO₂ litre^–1) around late-successional Alpine sedge communities of the Swiss Central Alps (2450 m) for four growing seasons (1992–1995) had no detectable effect on symbiotic N₂ fixation in Trifolium alpinum —the sole N₂-fixing plant species in these communities (74 ± 30 mg N m^–2 year^–1, A and E plots pooled).
2. This result is based on data collected in the fourth growing season showing that elevated CO₂ had no effect on Trifolium above-ground biomass (4·4 ± 1·7 g m^–2, A and E plots pooled, n = 24) or N content per unit land area (124 ± 51 mg N m^–2, A and E pooled), or on the percentage of N Trifolium derived from the atmosphere through symbiotic N₂ fixation (%Ndfa: 61·0 ± 4·1 across A and E plots) estimated using the ¹⁵N dilution method.
3. Thus, it appears that N inputs to this ecosystem via symbiotic N₂ fixation will not be dramatically affected in the foreseeable future even as atmospheric CO₂ continues to rise.     

DEVELOPMENT OF HEMOGLOBIN, HEMATOCRIT, AND ERYTHROCYTE VALUES IN GALÁPAGOS FUR SEALS

We studied the ontogeny of hemoglobin concentration, hematocrit and erythrocyte counts in the Galapagos fur seal ( Arctocephalus galapagoensis , Heller 1904). Two hundred and fifty-three animals were sampled between the ages of 22 d and > 8 yr, of which 46 were adult females. Body mass increased steadily with age from 6.1 ± 1.2 kg in 1-mo-old pups ( n = 27) to 28.5 ± 3.3 kg in adult females. Even adult females increased in mass with age. Hemoglobin (Hb), hematocrit (Hct), and red blood cell (RBC) values all increased in a logarithmic fashion with age up to 2 yr. Blood values for pups were Hct: 35.5 ± 4.1%; Hb: 12.9 ± 1.3 g/dl; RBC: 4.1 ± 0.3·10⁶/μl. Half-year-old fur seals (Hct: 42.1 ± 3.2%; Hb: 15.7 ± 1.3 g/dl; RBC: 4.9 ± 0.5·10⁶/μl; n = 50) were the oldest age group to show significantly lower blood values than adult females ( P < 0.001 for all three parameters). Yearling blood values (Hct: 47.2 ± 3.6%; Hb: 17.3 ± 1.6 g/dl; RBC: 5.6 ± 0.4·10⁶/μl; n = 56) did not differ significantly from those of adult females ( P < 0.32; P < 0.26; P < 0.23, respectively). Blood values of adult females were lower than those of 2-yr-olds (Hct: 49.6 ± 2.4%; Hb: 18.5 ± 1.2 g/dl; RBC: 5.7 ± 0.3·10⁶/μl; n = 31). These differences were significant only for RBCs ( P < 0.003). Up to the age of 1 yr, age was the best predictor for blood values, thereafter mass tended to be a better predictor. Female juveniles between the ages of 150 and 600 d had higher blood values than same-age males. The relationship of blood value development to diving activity is briefly described and the results are compared to values of other marine mammals. Ontogeny is discussed in relation to the development of these blood values in terrestrial mammals.     

Morphometry of some structural parameters affecting oxgyen diffusion in body trunk red muscle at different sizes of a tilapia, Oreochromis niloticus

Morphometric measurements were carried out on some of the structural parameters affecting oxygen diffusing capacity in red muscle of 15 specimens of O. niloticus body weight (b.w.) 0.65–812.3 g. Total capillary length and surface area and total morphometric oxygen diffusing capacity in body trunk red muscle had average values of 4792·7 (± 1740.7 s.e.) m, 415·4 (± 157·3 s.e.) cm² and 0·0213 (± 0·0075 s.e.) ml⁻¹ min⁻¹ cm⁻² mmHg respectively. When expressed as functions of b.w. these parameters had scaling values of 1·02, 1·07 and 0·993 respectively. These figures show a slight increase or almost no change in these structural parameters (which affect diffusion of oxygen to mitochondria in red muscle) per unit weight of fish. This should be important as the role of sustained swimming (by red muscle) becomes more important in larger tilapia. Oxygen diffusion distances were short [3·11 (±0·16 s.e.) μm] which facilitates diffusion of oxygen to mitochondria. The scaling value for oxygen diffusion distances of 0·067 (with respect to body weight) shows a slight increase in this parameter with development. This value is significantly different from zero.