Acute effects of the sodium salt of 2,4-D on the early developmental stages of bleak, Alburnm alburnus

The acute effects of a herbicide based on 2,4-D on embryos and larvae of bleak, Alburnus alburnus L. exposed to concentrations of 25–3200 mg 1⁻¹ for 12–48 hours were investigated. The 2,4-D delayed or stopped the development of embryos in their early stages, caused behavioural changes and morphological alterations. The L.C.₅₀ values for embryos varied between 12–9 and 15·9–4mg 1⁻¹ and for larvae from 51·6 to 111·2mg 1⁻¹ according to the time of exposure. The level of 2,4-D in Lake Balaton, or similar shallow lakes, without substantial injury of the littoral fish fauna should not surpass 0·5–1·0 mg 1⁻¹.     

Availability, uptake and turnover of glycerol in hypersaline environments

Abstract A sensitive assay for glycerol and other polyols was developed, based on periodate oxidation to formaldehyde, followed by a colorimetric assay with 3-methyl-2-benzothiazolone hydrazone. Apparent glycerol concentrations thus measured in saltern crystallizer ponds were around 20–36 μM, while in the Dead Sea, during a Dunaliella bloom, values were up to 27 μM. However, these values probably overestimate the glycerol concentrations present, as shown by labeled glycerol uptake experiments. Values of [K + S_n] (natural concentration + affinity constant) in saltern ponds were as low as 0.76–1.4 μM, with V_max values of 193–303 nmol 1⁻¹h⁻¹, and turnover times between 2.6–7.2 h at 35°C. Similar measurements in the Dead Sea were: [K + S_n] 0.07–1.41 μM, V_max values 160–426 nmol 1⁻¹h⁻¹, and turnover times in the range of 0.45–3.3 h.     

Biology of the acid-tolerant fish species Umbra pygmaea (De Kay, 1842)

In The Netherlands, the mudminnow Umbra pygmaea mainly occurs in water bodies with a pH ranging from 3–5 to 4–0, an alkalinity less than 0.1 meq 1⁻¹ and a calcium content below 100 μmol 1⁻¹. The abundance of mudminnows is inversely related to the pH and the presence of predatory fish species. Experimental studies show that If. pygmaea is extremely acid-tolerant. Exposure to media with pH values ranging from 4.0 to 7.0 did not cause mortality or significant changes in blood plasma osmolarity and haematocrit. At pH 3.0 no mortality was recorded although a strong decrease in the plasma osmolarity was observed. Exposure to pH 2.8 and calcium concentrations of 100 or 500 μ mol 1⁻¹ for 10 days caused 70% and 40% mortality respectively. The growth, reproduction and feeding of mudminnows are described. Age-growth and length-age relationships were calculated. Gonadal development started between September and January. The highest Gonado Somatic Index was recorded in spring. Spawning occurred from April to May. Fecundity of a sample of females was assessed. Nematocera provided the main item of diet of U. pygmaea; the microfauna ingested was dominated by Cladocera. The adaptations of mudminnows to acid environments are discussed.     

Some effects of low pH and calcium on the growth and tissue mineral content of yearling brown trout, Salmo trutta

Yearling brown trout, Salmo trutta , were exposed to low mineral content water (nominal concentrations of 20μmol 1⁻¹ magnesium, 7.7 μmol 1⁻¹ potassium, 44 μmol 1⁻¹ sodium) over a pH range of 4.0–5.2 with ambient calcium concentrations of 2.5–60 μmol 1⁻¹. All fish died at pH 4.0 and 4.2 irrespective of ambient calcium concentration and also at pH 4.4 with only 2–3 μmol 1 ⁻¹ calcium (that is calcium-free water except for that leached from the diet or excreted by the fish). Good growth rates were obtained over the remaining treatments which extended down to pH 4.4 with as little as 7 μmol 1⁻¹ calcium. When starved, weight loss was inversely correlated with pH. Effects on plasma chloride, percentage dry weight and calcium, potassium sodium, and phosphorus contents of skin, muscle and bone tissue were also investigated. These demonstrated pH effects on mineral metabolism in starved fish, but no effects were detected in fed fish.     

Effects of development, temperature and salinity on metabolism in eggs and yolk-sac larvae of milkfish, Chanos chanos (Forsskål)

Oxygen uptake rates and yolk-inclusive dry weiGhts were measured during the egg and yolk-sac larval stages of milkfish, Chanos chanos (Forsskal). Oxygen uptake by eggs and yolk-sac larvae was measured to assess the effects of four salinities (20,25,30,35 ppt) at 28°C. The effects of three temperatures (23,28,33°C) on oxygen uptake by yolk-sac larvae were determined at a salinity of 35 ppt. Dry weights were measured throughout embryonic development at 28°C and the yolk-sac stage at 23.28 and 33°C.
Oxygen uptake rates of eggs increased more than fivefold during embryogenesis (0.07±0.03 to 0.40 ± 03 μl O₂ egg ⁻¹ h ⁻¹;blastula to prehatch stage). Larval oxygen uptake did not change with age but was affected by rearing temperature (0.33 ± 0.08, 0.44 ± 0.07 and 0.63 ± 0.13 μl O₂ larva ⁻¹ h⁻¹ at 23, 28 and 33°C, respectively; Q₁₀= 1.93). Acute temperature changes from 28 to 33°C caused significant increases in oxygen uptake by embryos (Q ₁₀= 1.69–3.58) and yolk-sac larvae (Q ₁₀=2.55). Salinity did not affect metabolic rates.
Dry weight of eggs incubated at 28°C decreased 13% from fertilization to hatching. Incubation temperatures from 23–33°C did not affect dry weights at hatching. Rearing temperatures significantly affected the rate of larval yolk absorption (Q ₁₀= 2.25).     

Effects of river red gum, Eucalyptus camaldulensis, litter on golden perch, Macquaria ambigua

Aquaria with added river red gum, Eucalyptus camaldulensis , litter became hypoxic, with decreased pH and contained up to 30 mg 1⁻¹ tannin and lignin. Survival of golden perch, Macquaria ambigua , larvae in aquaria treated with a simulated annual litter density of 450 g m⁻² for 72 h was 14·9% for 15-day-old larvae and 0% for 8-day-old larvae. A litter density of 1223 g m⁻² resulted in total mortality for both age groups of larvae. Aeration increased survival of larvae to a minimum of 68·8% in 1223 g m⁻² litter treatments compared to 89·8% in aerated controls and 86·8% in non-aerated controls. A kinetic behavioural assay was used to detect alarm responses in golden perch larvae and juveniles exposed to leachates from river red gum bark, leaves and wood. Eight-day-old larvae exposed to bark and wood leachates (0·001–10 g 1⁻¹) exhibited an initial period of hyperactivity, followed by a concentration-dependent decrease in spontaneous activity. Larvae exposed to leaf leachates displayed only a decrease in spontaneous activity. Four-month-old juveniles exposed to wood leachates were also initially hyperactive, then progressively developed mild hypoactivity at increasing leachate concentrations. Juveniles exposed to wood leachates at 20g 1⁻¹ for 30min suffered 97·5% mortality in 96 h. Wood leachates induced dose-dependent lamellar fusion, epithelial dissociation and necrosis in the gills. The presence of toxic leachates and low oxygen availability in flooded river red gum forests may make these habitats unsuitable as nursery areas for native fish.     

Oxygen uptake in developing eggs and larvae of the cod, Gadus morhua L.

Unfertilised cod eggs showed a mean oxygen uptake rate at 5°C of 0.089 μl O₂, dry wt.⁻¹ h⁻¹; this gradually rose to 0.768 μl O₂ mg dry wt.⁻¹ h⁻¹ in eggs about to hatch. From hatching to complete yolk absorption larvae respired at 1.6 μl O₂, mg dry wt.⁻¹ h⁻¹. During starvation following yolk absorption, uptake fell significantly to 1.1 μl O₂, mg dry ⁻¹ h⁻¹. Much of this decrease in oxygen consumption was shown to be caused by reduction in activity. Loss of weight during the embryo and larval phases could not easily be reconciled with total oxygen consumption; it is suggested that cod embryos and larvae may not rely solely upon endogenous energy reserves during development.     

A light intensity threshold for schooling in the Atlantic mackerel, Scomber scombrus

The schooling behaviour of Atlantic mackerel was studied in a large tank at different light intensities in the range 12.6–1.8 × 10⁻¹⁰μEs⁻¹ m⁻². Variable light intensity was produced by accurately controlling the current to a green light-emitting diode (LED) 3 m above the experimental tank. Under high light levels (1.8 × 10⁻⁶μEs⁻¹ m⁻²) mackerel always formed a single school, whereas at lower levels (1.8 × 10⁻⁸μEs⁻¹ m⁻²) they swam as individuals. At light levels down to 1.0 × 10⁻⁶μEs⁻¹ m⁻² the mean nearest neighbour distance in a school remained relatively constant (0.3–0.9 body lengths), and individual mackerel swam along a path which deviated from the position of their nearest neighbours by less than 14°. As light dropped below 1.8 × 10⁻⁷μEs⁻¹ m⁻², both nearest neighbour distance and heading angle between nearest neighbours increased, with mean values of 1–1.8 body lengths and 23–92°, respectively, at 1.8 × 10⁻⁹μEs⁻¹ m⁻². The results are discussed in terms of ambient light conditions in the sea.     

Muscle buffering capacity of yellowtail fed diets supplemented with crystalline histidine

Juvenile yellowtail Seriola quinqueradiata (initial body mass of 22 g) were fed either a commercial diet (control, diet 1) or diets supplemented with histidine (diet 2), histidine+β-alanine (diet 3), or histidine+β-alanine+thyroxine (diet 4), for 6 weeks. The dietary treatment did not affect the final body mass. Free histidine levels of white muscle in the fish fed the diets supplemented with histidine (diets 2-4) were significantly higher (>62 mmol kg⁻¹ of wet tissue) than that of control group (42 mmol kg⁻¹ of wet tissue). Dietary supplementation of β-alanine (diet 3) or β-alanine+thyroxine (diet 4) failed to increase muscle anserine (β-alanyl-π-L.-histidine) level. Muscle buffering capacity of the range from pH 6·0 to 7·5 of the fish fed the diets 2-4 (41·6-42·7 mmol NaOH pH⁻¹ kg muscle⁻¹) reflected the increase of muscle histidine level, having slightly but significantly intensified compared to control fish (36·6 mmol NaOH pH⁻¹ kg muscle⁻¹). Most of the free amino acids other than histidine were significantly lower in the fish fed the diets 2-4 than in control fish. Thus, crystalline histidine supplemented to diets appears to be deposited in muscular tissue, and consequently enhance muscle buffering capacity in this species.     

Early life history of herring larvae in contrasting feeding environments determined by otolith microstructure analysis

Newly hatched autumn-spawned herring larvae Clupea harengus were released in two 2500-m³ outdoor mesocosms and reared over a 2-month period. Hydrographic conditions were similar in the two mesocosms, but the average plankton density was initially more than 10 times higher in mesocosm B compared to mesocosm A (>11⁻¹ v. <0.11⁻¹). Half-way through the experiment the feeding conditions reversed with three times higher average densities in mesocosm A than in mesocosm B (>31⁻¹ v. ∼11⁻¹). Herring larvae were sampled with a 0.3-m² two-chambered net twice weekly, and survivors were harvested by draining the mesocosms at the end of the experiment. Otolith growth trajectories of individual larvae were determined by relating radial otolith size with number of increments from the outer edge of the otolith (days before capture). The increment widths during the first 3 weeks after hatching, including the first-check size, were generally wider among larvae from mesocosm B (relatively good initial feeding conditions) than among those from mesocosm A (poor initial feeding conditions). The otolith growth pattern also confirmed that the surviving herring in mesocosm A belonged to the upper size range of larvae in the mesocosm after only 2–3 weeks from hatching; no such trend was found in mesocosm B. In both mesocosms the otolith size-at-age indicated that with the present sampling gear, herring larvae larger than 20–25 mm were underrepresented in the net samples. The information obtained from otolith-size-at-age is compared with other morphometric and biochemical measures of size and condition of larvae obtained throughout the experiment.     

ADSORPTION OF FULVIC ACID ON ALGAL SURFACES AND ITS EFFECT ON CARBON UPTAKE

Adsorption of Suwannee River fulvic acid (SRFA) to algal surfaces of three green algae was studied at environmentally relevant pH values (4 –7) and SRFA concentrations (5–100 mg·L ^{− 1}). The influence of adsorbed SRFA on carbon uptake of Scenedesmus subspicatus Chodat was also examined. Although no adsorption was observed at neutral pH values (pH 6 and 7), at pH 4 up to 31 mg SRFA·m ^{− 2} and at pH 5 up to 4 mg SRFA·m ^{− 2} was adsorbed to the algal surfaces. Electrophoretic mobility measurements of S. subspicatus demonstrated an increase in the negative surface charge of the alga in the presence of SRFA at pH 4. The adsorbed SRFA also influenced ¹⁴C uptake in S. subspicatus; in this case, enhanced carbon uptake could be related to the amount of adsorbed SRFA. The binding of humic substances by algal surfaces was interpreted as the result of hydrogen bonding and hydrophobic interactions.     

Embryonic and larval development of brown trout, Salmo trutta L.: exposure to trace metal mixtures in soft water

Freshly fertilized ova of brown trout, Salmo trutta L., were exposed to all possible mixtures of Al (6000 nmol 1¹), Cu (80 nmol 1⁻¹), Pb (50 nmol 1⁻¹) and Zn (300 nmol 1 ¹). In a separate experiment, newly hatched brown trout yolk-sac fry were exposed to Mn (1500 nmol 1⁻¹), Fe (2500 nmol 1 ¹), Ni (200 nmol 1⁻¹) or Cd (4 nmol 1 ¹), separately, and in mixtures with either Al or Cu. Both experiments were conducted in flowing, artificial softwater media nominally at pH 5.6 [Ca] 20 μmol 1 ¹ and 10° C.
Mortalities were high in fry subjected to treatments which contained both Al and Cu (31–72%), and to the Cu + Fe treatment (78%) compared with those from the other trace metal mixtures (0–22%). In all the treatments tested, fry exposed to trace metal mixtures containing Al and/or Cu had reduced whole body Ca, Na and K content, and seriously impaired skeletal calcification. Whole body Mg content was variable. In trace metal mixtures which contained Cu but not Al, the effects on fry survival and whole body mineral content were in general more deleterious than the corresponding mixtures but with Al present rather than Cu. The presence of Pb and/or Zn in mixtures with Al and/or Cu had a slight ameliorative effect in terms both of fry survival and whole body mineral content.     

Inhibition of enterobacteria and Listeria growth by lactic, acetic and formic acids

Minimum inhibitory concentrations (MIC) of undissociated lactic, acetic and formic acids were evaluated for 23 strains of enterobacteria and two of Listeria monocytogenes. The evaluation was performed aerobically and anaerobically in a liquid test system at pH intervals of between 4.2 and 5.4. Growth of the enterobacteria was inhibited at 2–11 mmol 1⁻¹, 0.5–14 mmol 1⁻¹ and 0.1–1.5 mmol 1⁻¹ of undissociated lactic, acetic and formic acids, respectively. The MIC value was slightly lower with anaerobic conditions compared with aerobic conditions. The influence of protons on the inhibition was observed for acetic acid at the low pH values. Undissociated lactic acid was 2 to 5 times more efficient in inhibiting L. monocytogenes than enterobacteria. Acetic acid had a similar inhibitory action on L. monocytogenes compared with enterobacteria. Inorganic acid (HCl) inhibited most enterobacteria at pH 4.0; some strains, however, were able to initiate growth to pH 3.8. The results indicate that the values of undissociated acid which occur in a silage of pH 4.1–4.5 are about 10–100 times higher than required in order to protect the forage from the growth of enterobacteria and L. monocytogenes.     

Larval competition for food between wild and hatchery-reared ayu, Plecoglossm altivelis Temminck et Schlegel, in culture ponds

To examine the larval competition between wild and hatchery ayu in the culture ponds, mixed rearing of 580 000 wild and 520 000 hatchery larvae was carried out in two 25-m³ ponds for 3 months, in contrast to the control in which 860 000 wild larvae were reared in another pond.
The number of wild larvae in the mixed-rearing treatments decreased rapidly 20 days after the start of mixed rearing, in contrast to hatchery larvae. Mortality of wild larvae was almost 100% at the end of the experiment (3 months), while the hatchery larvae showed the usual survival rate of 15–16%. In the control pond, however, 16% of the wild larvae survived. The wild larvae grew much slower (0.10mmday ^-1) than the hatchery larvae (0·26 mm day ^-1) in the mixed-rearing ponds, whereas the wild larvae in the control pond showed almost the same growth rate (0·24 mm day ^-1) as hatchery larvae. On day 6 the gut fullness of wild larvae was only 30% of that of the hatchery larvae in the mixed-rearing experiments. On day 46 the wild larvae occurred deeper in the mixed-rearing ponds than the hatchery larvae. This depth difference in vertical distribution appeared to cause a disadvantage for the wild larvae in the competition with hatchery larvae, since the food was supplied at the surface. Thus, the wild larvae starved and died.     

EFFECTS OF CADMIUM TOXICITY ON GROWTH AND ELEMENTAL COMPOSITION OF MARINE PHYTOPLANKTON

Some classes of marine phytoplankton are believed to be more tolerant of high concentrations of trace metals than others, but the results of experimental tests of this hypothesis are ambiguous. Eleven species of phytoplankton representing five classes were grown in Aquil medium containing Cd concentrations between 10⁻⁸ and 10⁻⁵ M ([Cd²⁺]= 10^−9.85 to 10^−6.84 M), and growth rates and intracellular concentrations of Cd, C, N, and S were measured. The mean Cd²⁺ concentration (pCd⁵⁰) that reduced the growth rate of each species to 50% of its maximum varied by 2.5 orders of magnitude, from 10^−6.23 for Emiliania huxleyi to 10^−8.79 for Synechococcus sp. Taxonomic trends in Cd resistance were not apparent in these data. Cadmium quotas (mol Cd·L⁻¹ cell volume) were lowest in species of Bacillariophyceae (ANOVA, P < 0.001), suggesting that they might regulate Cd transport differently than other taxa. Cellular S:C molar ratios increased in four of seven phytoplankton grown at high pCd (7.37–6.84) compared to low Cd ion concentrations (no added Cd), a result of increases in S·L⁻¹ cell volume. Nitrogen:carbon molar ratios were also higher in Cd-exposed phytoplankton, as changes in N and S were highly correlated ( r = 0.98, P < 0.0001). In two species that were examined, S:C ratios increased as a linear function of increasing Cd concentration. The results demonstrate large variability in Cd resistance among phytoplankton that is primarily a function of interspecific differences in Cd detoxification.     

Photosynthetic characteristics of the submerged macrophyte Ceratophyllum demersum

The photosynthetic and growth characteristics of Ceratophyllum demersum L. were investigated under laboratory conditions which simulated those encountered in the plants' normal environment. The carbon fixation rate of C. demersum measured with ¹⁴C at light and carbon saturation at pH 8.0 was 4.48 mg C (g ash-free dry weight)⁻¹ h⁻¹. It was lower at pH 6.5 than at pH 8.0. The light use efficiencies in quiescent plants and actively growing plants were 6.3 and 8.7 × 10⁻⁹ kg CO₂ J⁻¹, respectively, with corresponding maximum photosynthetic rates of 2.67 and 4.36 mg C (g ash-free dry weight)⁻¹ h⁻¹. Photorespiration in actively growing plants consumed 24% of the carbon fixed. Incubation with DCMU demonstrated that about one-third was refixed. The optimum temperature for carbon fixation was 25°C. The C₃-photosynthetic pathway was the main operational route as indicated by the early photosynthetic products (largely C₃-acids) and the absence of Krantz anatomy and the chlorophyll a:b ratio (2.7). The maximum relative growth rates ranged from 0.025 to 0.041 g ash-free dry weight (g ash-free dry weight)⁻¹ day⁻¹ in the field (Lake Vechten, 1 to 3 m depth classes).     

Cultural conditions for production of glucoamylase from Lactobacillus amylovorus ATCC 33621

Lactobacillus amylovorus ATCC 33621 is an actively amylolytic bacterial strain which produces a cell-bound glucoamylase (EC 3.2.1.3). Conditions of growth and glucoamylase production were investigated using dextrose-free de Man-Rogosa-Sharpe (MRS) medium in a 1.5 I fermenter, with varying dextrin concentration (0.1–1.5% (w/v)), pH (4.5–6.5) and temperature (25–55°C). Cell extracts were prepared by subjecting cells to treatment with a French Pressure cell in order to release intracellular proteins. Glucoamylase activity was then assayed. The effects of pH (4.0–9.0), temperature (15–85°C) and substrate (dextrin and starch, 0–2% w/v) concentration on crude enzyme activity were investigated. Optimal growth was obtained in MRS medium containing 1% (w/v) dextrin, at pH 5.5 and 37°C. Glucoamylase production was maximal at the late logarithmic phase of growth, during 16–18 h. Crude enzyme had a pH optimum of 6.0 and temperature optimum of 60°C. With starch as the substrate, maximal activity was obtained at a concentration of 1.5% (w/v). The effects of ions and inhibitors on glucoamylase activity were also investigated. Enzyme activity was not significantly influenced by Ca²⁺ and EDTA at 1 mmol 1⁻¹ concentration; however Pb²⁺ and Co²⁺ were found to inhibit the activity at concentrations of 1 mmol 1⁻¹. The crude enzyme was found to be thermolabile when glucoamylase activity decreased after about 10 min exposure at 60°C. This property can be exploited in the brewing of low calorie beers where only mild pasteurization treatments are used to inactivate enzymes. The elimination of residual enzyme effect would prevent further maltodextrin degradation and sweetening during long-term storage, thus helping to stabilize the flavour of beer.     

Resistance to anoxia of Chironomus plumosus and Chironomus anthracinus (Diptera) larvae

Survival of the midge larvae Chironomus plumosus and C. anthracinus in anoxia at 4°C was investigated. C. plumosus survived about twice as long as C. anthracinus . The corresponding LT 50 values were ca 205 and 100 d. There was no statistically significant difference between the survival in anoxia and in aerated water, which indicates that the main reason for death in anoxia is not the absence of oxygen. This main reason is presumed to be starvation. The presence of undissociated H₂S in low concentrations (ca 1.2 mg 1⁻¹) did not influence the survival. In anoxia the larvae were usually motionless and did not feed but they increased in weight due to uptake of water. During 43 d C. plumosus and C. anthracinus increased ca. 10%.     

A limnological investigation of the tarn Syslaktjønn, W Norway

An investigation covering hydrography, chemistry, vascular and cryptogamic plants, nitrogen fixation, phytoplankton biomass and production, and zooplankton was carried out from April to November 1976 in tarn in W Norway, The volume of the tarn was 18000 m³ and the turnover time about 30 d. Temperature ranged between 3.6 and 23.4°C and pH between 4.8 and 5.5. Nuphar luteum and Carex rostrata were the two dominating vasculars-with biomasses of 117 and 97 g m⁻², respectively The biomass of the bryophytes ( Sphagnum spp.) was about 510 g m⁻² and the production of the order 0.2–2.1 μg (mg d.w.)⁻¹h⁻¹. Nitrogen fixation in association with Sphagnum spp. was estimated at 25 g yr⁻¹ for the whole tarn. Phytoplankton was dominated by diatoms, green algae and chrysophyceans. The chlorophyll a content ranged from 2 to 20 mg m⁻³ and the carbon assimilation rates from 0.03 to 20 mg C m⁻³h⁻¹ at 0–4 m depths. Production in the period was of the magnitude 22 g C m⁻². The copepod Eudiaptomus gracilis was the most common netzooplankter. Large numbers of rotationrians were found during summer.     

Growth response of pikeperch larvae in relation to body size and zooplankton abundance

The most critical period at onset of feeding in pikeperch Stizostedion lucioperca is short (<5 days at 20° C). The larvae are sensitive to prey density during the first week of exogenous feeding. First-feeding larvae of 6.5 mm total length ( L_T ) needed prey densities of >585 prey l⁻¹ to maintain mass (C_maint), whereas 5 days older larvae of 7 mm L _T C_maint= 55 prey l⁻¹ and for 11 mm L _T larvae C_maint<10 prey 1⁻¹. Changes in specific growth rates for larvae of 7 and 11 mm were similar to a type-II functional response curve reaching a specific growth rate of 26 and 30% day⁻¹, respectively, at 1000 prey l⁻¹, whereas the 6·5 mm larvae showed a linear growth response reaching a specific growth rate of only 9% day ⁻¹ at 1000 prey l⁻¹. The results suggest that prey density is a limiting factor, which might contribute to the high variation between year-class Strengths.