Evaluation of flow cytometry as a method for quantification of circulating blood cell populations in salmonid fish

Flow cytometry was used to estimate the proportions of different blood cell types in brown and rainbow trout. On the basis of forward light scatter and 90° side scatter three populations were differentiated. The relative abundance of these cells correlated with that of erythrocytc (r²= 0.994), lymphocyte plus thrombocyte(r²= 0.676) and neutrophil populations (r²= 0.571) enumerated by direct microscopy. By density gradient separation of cells, cell sorting and acridine orange staining it was confirmed that these cell types could be assigned to the populations detected. Changes in blood cell populations were monitored by flow cytometry in a group of experimental fish placed under confinement stress. Flow cytometry proved to be a rapid and reliable method for monitoring cell population dynamics in fish blood.     

Groundwater controls on nutrient cycling in a Mojave desert stream

1. Groundwater fluxes of nitrogen and dissolved organic carbon (DOC) were investigated in Grape Vine Canyon Stream in the Mojave Desert focusing on the rate of inputs and the fate of groundwater-derived nutrients in the stream. Discharge rates from different ground waters were measured using an end-member mixing model coupled with injections of a conservative solute tracer into the stream channel.
2. In surface water, nitrate concentration averaged 1.13 mg N L^–1 and DOC concentration averaged 1.82 mg C L^–1.
3. Groundwater discharge into Grape Vine Canyon Stream was derived from three sources. Nitrate concentration varied among the three groundwater sources with mean concentrations of 0.56, 0.94 and 0.08 mg N L^–1. DOC, in contrast, did not vary among ground water sources, with an overall average concentration of 2.96 mg C L^–1.
4. In the surface stream, nitrate concentration was two-fold greater than the concentration predicted from groundwater input, indicating that in-stream processes generated nitrate. Stream DOC concentration was lower than predicted based upon groundwater input rate. The production of nitrate and loss of DOC suggest that DOC is lost through mineralisation of dissolved organic matter, possibly resulting in the mineralisation of dissolved organic nitrogen to ammonium and subsequent transformation to nitrate via nitrification. In further support of this hypothesised linkage, DOC loss explained 80–89% of the variance in nitrate production in Grape Vine Canyon Stream.     

Effect of competition and leaf age on visible and infrared reflectance in pine foliage

Abstract. The effect of plant competition on spectral reflectance in the 400–2500 nm wavelength region was determined for 3-month-old and 15-month-old leaves of loblolly pine ( Pinus taeda L.). Strong competition decreased water potentials and Mg concentrations, and increased K in young and old leaves. Also, competition decreased Ca and total chlorophyll in young leaves. As measured with a scanning radiometer, reflectance in young leaves at 551 nm decreased from 20 to 14% as water potentials increased from -2.2 to -0.9 MPa (r²= 0.82). For young and old leaves reflectance at 551 nm decreased from 20 to 10% with increasing total chlorophyll (r²= 0.64). Reflectance decreased slightly with increasing K in young leaves (401 nm, r²= 0.55), and with increasing Mg in old leaves (470 nm, r²= 0.57). Increased visible reflectance under strong competition may have resulted primarily from decreased water potentials. Reflectances were much greater in young versus old leaves in the 750–1300 nm range, and were greater in old rather than young leaves from 1400–2500 nm. Infrared reflectances were not, however, significantly affected by competition.     

Effect of temperature on swimming performance of sea bass juveniles

At four temperatures ( T= 15, 20, 25 and 28° C) swimming performance of Dicentrarchus labrax was significantly correlated with total length (23–43 mm L _T); r²=0.623–0.829). The relative critical swimming speed ( RU _crit= U _crit L _T⁻¹), where U _crit is the critical swimming speed, was constant throughout the L _T range studied. The significant effect of temperature on the relative critical swimming speed was described binomially: RU _crit=−0.0323T²+ 1.578 T −10.588 (r²=1). The estimated maximum RU _crit (8.69 L _T s⁻¹) was achieved at 24.4° C, and the 90% performance level was estimated between 19.3 and 29.6° C.     

Life cycle, production, microdistribution and diet of the damselfly Euphaea decorata (Odonata: Euphaeidae) in a Hong Kong forest stream

Euphaea decorata in Tai Po Kau Forest Stream (Hong Kong) was univoltine. Most recruitment took place in summer, and larval growth proceeded throughout the year. Life-cycles recorded in 1977–78 and 1978–79 were similar. Annual production estimates, using the removal-summation, instantaneous growth and size-frequency methods, were more similar for the 1978–79 generation (ranging from 158.7–174.7; mean 1671 mg dry wt m^-2) than for the 1977–78 generation (93.9-173-6; mean 131.7 mg dry wt m^-2). Mean biomass was similar for both generations (ranging from 33.5–33.9 mg dry wt m^-2), and mean P/B ratios were 3.9 1977–78) and 5.0 (1978–79). These are the first estimates of annual production by an Oriental stream insect.
Larvae were most abundant at microsites in the middle of the stream. Multiple regression analysis indicated that substratum characteristics were a major determinant of microdistribution. Euphaea decorata apparently favoured poorly-sorted sediments with highly peaked grain size-frequency distributions, containing few fine particles.
The carnivorous larvae showed ontogenetic changes in diet. Small individuals consumed mainly chironomid (Diptera) larvae; the diet expanded to include (successively) larvae of Ephemeroptera and Trichoptera as E. decorata grew. Seasonal changes in diet were also apparent, although larval diets during spring and summer were similar. There was also considerable overlap between autumn and winter diets. Ontogenetic influences upon prey consumed were not sufficient to account for the observed seasonal differences.     

Nutrient and energy content of freshwater fishes

The analyses of fresh total body homogenates of 2385 freshwater fishes from 17 species originating in natural waters and aquaculture facilities resulted in average values of 19.1–37.8% dry matter, 13.6–18.3% crude protein, 0.8–18.3% crude fat, 0.16–2.17% nitrogen-free extracts (NFE), 2.2–2.9% nitrogen and 0.39–1.02% phosphorus. The gross energy content ranged from 3.83 to 11.49 MJ/kg. With increasing fat content in the fishes, water content decreased and dry matter content increased. Through this interdependence, a significant correlation was found between dry matter and crude fat and energy content (r²=0.92–0.97). The values of crude protein, crude ash, NFE, nitrogen and phosphorus did not show any significant influence on dry matter content (r²=0.0005–0.46). Based on the significant correlation between dry matter and gross energy (gross energy [MJ/kg]=0.0253 dry matter [%]^1.6783), the energy content of freshwater fishes can easily be estimated by the dry matter content in the total body.     

An application of the plaster dissolution method for quantifying water velocity in the shallow hyporheic zone of an Appalachian stream system

1. A method for quantifying interstitial water velocity based on the dissolution rate of plaster of Paris standards was developed as part of a study of vertical, longitudinal (1–4 order sites) and seasonal variation in the biotic and physical characteristics of the shallow hyporheic zone (0–30 cm) of a headwater stream system in West Virginia, U.S.A.
2. A calibration model was developed using a water velocity simulation tank to relate mass loss of plaster standards to water velocity and temperature. The model was then used to calculate water velocity through artificial substrata embedded in the shallow hyporheic zone of four stream reaches based on in situ mass loss of plaster standards.
3. Water velocity in the hyporheic zone increased with stream order, was highest in early spring and winter during high stream base flows, and decreased with depth into the substratum. There was a strong interaction between depth and season: during periods of high stream discharge, water velocity through the upper level of the shallow hyporheic zone (0–10 cm into the substrate) increased disproportionately more than velocity at greater depths. Mean interstitial velocity in March ranged from 0 cm s^–1 in the lowest level (20–30 cm) to 3.5 cm s^–1 at the upper level (0–10 cm) at the first‐order site, and from 2.5 cm s^–1 (20–30 cm) to 9.5 cm s^–1 (0–10 cm s^–1) at the fourth‐order site. Gradients in stream discharge and sediment permeability accounted for treatment effects.
4. Use of calibrated data improved the ability to resolve among‐season differences in interstitial water movement over the use of uncalibrated mass loss data. For some applications of the plaster standard method, empirical calibration may not be necessary.     

Vertical distribution and abundance of invertebrates within the sandy substrate of a low-gradient headwater stream

SUMMARY. 1 The vertical distribution of invertebrates (>0.053 mm) was studied in a sandy-bottomed, first-order stream on the Coastal Plain of Virginia, U.S.A. Invertebrate species composition, abundance and bio-mass were determined monthly over one year at sediment depth intervals of 0–1, 1–5, 5–15,15–30 and 30–40 cm.
2. The subsurface community was numerically dominated by species of Chironomidae, Nematoda and Crustacea, while much of the biomass was due to early instars of several species of Trichoptera.
3. Invertebrate density and biomass decreased significantly with depth in the substrate (ANOVA; P <0.05). Annual mean density decreased from 1,346,844 individuals m⁻³ at the surface to 13,578 individuals m⁻³ at 15–30 cm. Annual mean biomass decreased from 66.30 g m⁻³ at the surface to 0.44 g m⁻³ at 15–30 cm.
4. Dissolved oxygen decreased markedly from the surface to the 5 cm depth in the substrate, anaerobic conditions often occurring below 10 cm. Density and biomass both showed a significant positive relationship with dissolved oxygen concentration (Linear regression; P <0.05).
5. Physical forces were important in structuring the subsurface invertebrate community. Besides low dissolved oxygen concentration, sediment scouring resulting from storm discharge dramatically reduced density and biomass     

The role of life history in the relationship between population dynamics and environmental variability in two Mediterranean stream fishes

Chub Squalius torgalensis and nase Chondrostoma lusitanicum , in a Mediterranean stream, showed important differences in life-history traits and population dynamics. Both species reached mean maturity at age 2 years. Chub lived up to age 5 years, spawned in March to June, grew at a maximum rate of 0·59 mm mm⁻¹ year⁻¹ and showed a low reproductive allocation, with fecundity and egg size increasing with body size. Nase lived up to age 4 years, spawned in January to April, grew at a maximum rate of 0·46 mm mm⁻¹ year⁻¹ and showed a high reproductive allocation, with egg size independent of body size. Both chub and nase showed moderate fluctuations in population size during 1991–1998, but differed in factors driving density at age. Density of age 1 year juvenile chub decreased following severe summer droughts and proportionate survival prevailed thereafter. Density of age 2 year adult nase decreased following severe spring floods, but neither environmental nor parental stock effects were detected for juveniles and older fishes. The results illustrated the interplay between life history and environmental variability in driving fish population dynamics, with impacts of both summer droughts and spring floods being contingent on species-specific patterns of spawning and reproductive investment.     

The production of brown trout, Salmo trutta in tributaries of the Upper Wye, Wales

The population density, age structure, biomass, growth and production of brown trout were investigated in four tributaries of the upper River Wye. The populations at each site were largely maintained by immigration from nursery areas. Abundance of separate year classes at sites on the three largest tributaries reached a peak at age 2+. On the smallest stream numbers reached a peak at 1+. Recruitment occurred throughout the year but decreased with age of year classes. Maximum O+ densities ranged from 0.04 to 0.89 m⁻², and >0+ densities from 0.13 to 0.59 m⁻². Average total biomass in 1975 ranged from 2.6 to 14.2 g m⁻². Within the study sites annual trout production in 1975 ranged from 2.9–19.7 g m⁻². Production values were dependent on age structure and population mobility at the study sites. In the three largest streams 2+ and 3+ fish contributed 66.3–88.3% of total production whilst 1+ and 2+ fish contributed 74.5–84.5 % of the total in the smallest stream. The mobile (non-resident) component of the population accounted for up to 60–70% of production at certain times of the year, but over the year (1976) accounted for =30 % of total production. The resident component of the highest annual production value (19.7 g m⁻²) was estimated to be between 15.0 and 18.2 g m⁻².     

Concentrations and annual loading of dissolved organic matter in a small moorland stream

SUMMARY. 1. Dissolved organic matter (DOM) was measured at 8h intervals over a 1 year period in a stream draining 51 ha of moorland with peaty soils.
2. DOM concentrations increased with increasing stream discharge from low flow values of 0–3 mg 1⁻¹ to maximum values of 30 mg 1⁻¹. There were also seasonal differences of up to 13mgl⁻¹ between August (maximum) and February, and differences of about 2.5 mg l⁻¹ between rising and falling stage samples.
3. Seasonal variation was closely related to mean temperature.
4. Total loss over the year was 168 kg ha⁻¹ DOM (84 kg ha⁻¹ C), larger than previous estimates for upland sites.     

Energy and ration requirements of juvenile Pacific halibut (Hippoglossus stenolepis) based on energy consumption and growth rates

Growth of captive juvenile Pacific halibut was linearly related to energy consumption (J g⁻¹ day⁻¹) at 4°C by the following equation: growth (% body weight (b.w.) day⁻¹)=0–007 (consumption J g⁻¹ day⁻¹)– 0.192; r² =0.81. Weight gain was independent of size for fish between 9 and 7000 g when growth was expressed as a function of consumption in J g⁻¹ day⁻¹. Maintenance ration determined in feeding–growth experiments averaged 27.4 J g⁻¹ day⁻¹ at 4–0°C. Small halibut ate significantly more food than large fish. Single meals following 2 day fasts averaged 4.1% b.w. for halibut under 100 g, 1.72% b.w. for 1.2 kg fish and 1.1% B.W. for 6.8 kg fish. Both large and small size categories of halibut tended to evacuate their meal in about 3 days even though small fish ate relatively larger meals. Minimum estimates for daily ration to achieve growth rates observed in the Gulf of Alaska were approximately 0.5 to 2.4% b.w. day⁻¹ depending on fish size and whether northern shrimp or yellowfin sole were their prey.     

The relative abundance of brown trout in acidic softwater lakes in relation to water quality in tributary streams

The influence of the water quality of tributary streams on the relative abundance in benthic gillnet catches (catch per unit effort, cpue) of allopatric brown Salmo trutta was assessed in associated acidic, softwater lakes. The study was carried out over 6 years (1989–1994) in 15 lakes located at altitudes between 230–715 m a.s.l. in two Norwegian catchments. The water quality of the main inlets and outlets varied little, as indicated by their of pH range (4·93–5·51) and calcium concentrations (0·19–0·44 mg 1⁻¹), but varied more with respect to concentrations of inorganic, monomeric aluminium (7·0–41·0 μg l⁻¹). Most of the lakes were also fed by secondary streams with better water quality: a maximum pH of 6·56, calcium levels of up to 0·74mg 1⁻¹, and inorganic aluminium levels as low as l·0 μg 1⁻¹. The cpue was inversely correlated with lake altitude ( r ²=0·50), and thus was adjusted to a mean altitude. The calcium concentration in the richest secondary stream to each lake, its richness judged on the basis of its acid-neutralizing capacity, had the highest predictive power of the variability in cpue ( r ² = 0·49).The calcium content in the other secondary streams or in the main inlets and outlets did not correlate with cpue. Alkalinity in the main outlets correlated to some extent with cpue ( r ² = 0·27). It is suggested that secondary streams with good water quality provide important refuges for the recruitment of brown trout in acidic softwater lakes.     

Micro-environmental characteristics of filamentous algal communities in flowing freshwaters

SUMMARY. 1. Filamentous algae in flowing freshwaters can represent a spatially and temporally distinct sub-habitat for epiphytic diatom communities. This sub-habitat is described in a low discharge, spring-fed stream with extensive filamentous green algal mats, and in a tuft of Cladophora glomerata (L.) Kützing from a large river.
2. Oxygen micro-electrodes, a thermistor current velocity probe, a standard pH probe and water chemistry were employed to assess spatial heterogeneity. Temporal patterns of epiphyte colonization were evaluated on filamentous artificial substrates.
3. There were steep spatial gradients in the low discharge stream. At mid-day, O₂ ranged from 0–1.5 times air-saturated O₂ concentrations, pH varied from 7.25–8.0, and current velocity spanned 0–0.5 m s⁻¹. Areas near the surface of algal mats had high O₂, pH and current velocity. These patterns were correlated with epiphyte community structure.
4. In the interior of C. glomerata tufts O₂ concentration was raised and current velocity depressed compared to the surrounding water, even when external current velocity was as high as 0.4 m s⁻¹.
5. After thirty-five days of colonization of artificial substrate in the low discharge environment, epiphyte communities were similar to those on filamentous atgae. Epiphyte diversity on artificial substrates subsequently decreased compared to natural substrates as did the similarity between the types on substrates, suggesting that microscale renewal of epiphyte habitat (growth of filamentous algae) maintains high epiphytic diversity.     

Population dynamics in a cyclic environment: consequences of cyclic food abundance on tawny owl reproduction and survival

1. Understanding which factors regulate population dynamics may help us to understand how a population would respond to environmental change, and why some populations are declining.
2. In southern Finland, vole abundance shows a three-phased cycle of low, increase and decrease phases, but these have been fading out in recent years. During five such cycles (1981–1995), all tawny owls Strix aluco were censused in a 250-km² study area, and their reproduction and survival were monitored.
3. Males and females showed similar dynamics, but experienced breeders recruited more offspring and had higher survival than first breeders. Offspring recruitment, but not survival of breeding individuals varied in accordance with vole abundance.
4. The population's numerical response to prey abundance was primarily due to first-breeding individuals entering the population in the increase phase when immigration was the highest. First-breeding birds were younger, but experienced breeders were older in more favourable vole years.
5. A stage-specific matrix population model integrating survival and fecundity showed that, despite obvious variation in fecundity between vole cycle phases, this variation had limited importance for overall tawny owl population dynamics, but that the survival of experienced breeders during the low phase is most important for population growth.
6. Model and data agreed that the vole cycle drives the dynamics of this avian predator by limiting the recruitment of new breeders during the low phase. Population dynamics hence differ not only from the classic example of the species in a more temperate region in the UK where the number of territories is stable across years, but also from the dynamics of other avian vole predators in Fennoscandia where the recurring crash in vole abundance drastically lowers adult survival thereby creating vacancies.     

Temporal patterns of milk production in Antarctic fur seals (Arctocephalus gazella)

The timing of milk production in Antarctic fur seals was studied at Bird Island, South Georgia. Like all lactating otariid seals (Pinnipedia: Otariidae), Antarctic fur seal females alternate between short nursing periods ashore and regular foraging trips to sea. Females do not necessarily return to the colony with full mammae, which indicates that mammary volume capacity is unlikely to limit foraging trip duration. Upon arrival at the colony, milk fat (r²= 0.33, P < 0.04) and protein (r²= 0.60, P < 0.002) content were positively correlated to the time spent at sea. A similar trend was observed in the milk produced on land. The rate of milk energy production was much lower at sea (5.02 ± 0.05 MJ day^-1) than on land (23.66 ± 4.4 MJ^-1 day^-1). The rate of milk energy production during the foraging trip was negatively correlated to the time spent at sea (r²= 0.29, P < 0.05), whereas the rate of milk energy production on land was positively correlated (r²= 0.61, P < 0.001) to the duration of the preceding foraging trip. The total amount of milk energy delivered to the pup during each twoday nursing period was positively correlated (r²= 0.60, P < 0.002) to the duration of the previous foraging trip. The overall rate of milk energy delivery, however, was independent of foraging trip duration. This accords with previous observations that the growth rates of Antarctic fur seal pups are unaffected by maternal foraging trip duration patterns.     

The relationship between foraging behaviour and energy expenditure in Antarctic fur seals

By using time-depth recorders to measure diving activity and the doubly-labelled water method to determine energy expenditure, the relationship between foraging behaviour and energy expenditure was investigated in nine Antarctic fur seal females rearing pups. At-sea metabolic rate (MR) (mean of 6.34 ± 0.4 W. kg^-1; 4.6 times predicted BMR) was positively correlated to foraging trip duration (mean of 4.21 ± 0.54 days; r²= 0.5, P < 0.04). There were no relationships between MR and the total number of dives, the total time spent diving or the total vertical distance travelled during the foraging trip. There was, however, a close negative sigmoidal relationship (r²= 0.93) between at-sea MR and the proportion of time at sea spent diving. This measure of diving behaviour may provide a useful, inexpensive means of estimating foraging energy expenditure in this species and possibly in other otariids. The rate of diving (m.h^-1) was also negatively related to at-sea MR (r²= 0.69, P < 0.005). Body mass gain during a foraging trip had a positive relationship to the time spent at sea (r²= 0.58, P < 0.02) and the total amount of energy expended while at sea (r²= 0.72, P < 0.004) such that, while females undertaking long trips have higher metabolic rates, the energetic efficiency with which females gain mass is independent of the time spent at sea. Therefore, within the range of conditions observed, there is no apparent energetic advantage for females in undertaking foraging trips of any particular duration.     

Biomass and oxygen dynamics of the epiphyte community in a Danish lowland stream

SUMMARY. 1. We examined the abundance and oxygen metabolism of epiphytic organisms on the dominant macrophyte, Potamogeton pectinatus , in headwaters of the eutrophic River Suså. Microbenthic algae were abundant in the stream during spring and macrophytes during summer.
2. The low macrophyte biomass in spring supported a dense epiphyte cover whereas the high macrophyte biomass during summer had a thin epiphyte cover of 10–100-fold lower abundance per unit area of macrophyte surface. The epiphyte community was dominated by microalgae in spring and by heterotrophs, probably bacteria, during summer. This seasonal shift was shown by pronounced reductions of the chlorophyll a content (from 2–3% to 0.1–0.7% of organic DW), the gross photosynthetic rate (from 20–85 to 3–15 mg O₂, g-¹ organic DW h⁻¹) and the ratio of gross photosynthesis to dark respiration in the epiphyte community (from 5–18 to 1). The reduced contributions of epiphytic microalgae correlated with reduced light availability during summer.
3. Both the density and the photosynthetic activity of epiphytic algae were low on a stream area basis relative to those of microbenthic algae and macrophytes. Rapid variations in water velocity and extensive light attenuation in water and macrophyte stands probably constrained the development of epiphytic algae. The epiphyte community was more important in overall stream respiration, contributing c. 10% to total summer respiration and c. 20% to summer respiration within the predominantly heterotrophic microbial communities on sediments and macrophyte surfaces.     

The populations of fish in the River Tees system on the Moor House National Nature Reserve, Westmorland

The age, growth, population densities and annual production of an isolated population of brown trout ( Salmo trutta L.) and bullhead ( Coitus gobio L.) were investigated over a 5 year period in Trout Beck in the northern Pennines. Additional data were obtained from two other sites; Great Dodgen Pot Sike and the River Tees.
Trout growth rates were low, particularly in Dodgen Pot Sike, but ages up to VIII were recorded. Bullhead growth rates were similar to those in the Tees at Cow Green (Crisp et al. , 1974), though the Moor House bullheads had a higher survival rate. Population densities of trout ranged from 0.10–0.22/m², with a fry contribution of 0.80%. The bullhead population in Trout Beck was sparse, generally 0.1/m², though an 0 group value of 0.46/m^a was estimated in 1967. At Tees Bridge values ranged from 2.5–7/8/m². Most trout matured by age III, though in Great Dodgen Pot Sike only half of the females were mature at age IV.
Annual production was estimated from Allen graphs. Trout production ranged from 1.02–3.50 g/m²/year, the fry contributing up to 51.5% of the total. A value of 0.48 g/m²/year was obtained for bullheads in the Trout Beck system in a year of good recruitment, whilst bullhead production at Tees Bridge was 7.43 g/m²/year, with fry and I group contributing 48.7 and 37.5% respectively. The high survival rate, irregular recruitment, and poor growth of these high altitude populations is discussed. Growth and reproductive peculiarities of the Great Dodgen Pot Sike trout are also discussed.     

Phenology of Sirodotia suecica (Batrachospermaceae, Rhodophyta) in a high-altitude stream in central Mexico

The morphology and phenology of a monoecious population of Sirodotia suecica Kylin was evaluated seasonally in a central Mexican fifth-order high-altitude stream. Abundance of gametophytes is positively correlated with concentrations of total dissolved solids, specific conductivity and total ionic concentration; being present the whole dry season and disappearing in the rainy season. The gametophytes grew in eutrophic circumstances and particular microhabitat conditions: high current velocity (66–122 cm s⁻¹), low irradiance (75–263 µmol photons m⁻² s⁻¹) and shallow depth (7–26 cm). Percent cover of gametophytes ranged from 5–90% and significant differences in abundance were not observed when alga was present. Some morphological and reproductive characteristics seem to be adaptations to high current velocity: abundant secondary branches, spermatangia and carpogonia. 'Chantransia' stage, microscopic creeping filaments associated with the base of the gametophyte, were not observed in natural conditions. In terms of reproductive success, the population studied can be regarded as highly efficient, considering the high fertilized carpogonia rate, similar to monoecious populations in lotic habitats. However S. suecica was not common in the study region because it was restricted to particular microhabitat conditions.