Ovarian contraction in various teleost families

Fishes from 12 species (a minimum of three fish of each species), representing nine families were examined for ovarian contraction. An ovary from each fish was removed, placed in a muscle bath and assessed for contractions using a force-displacement transducer. After at least 30 min of observation for spontaneous contractions, acetylcholine (ACh) was added to the muscle bath to assess the ovary's capacity for contraction. At least one third of the ovaries from each species contracted spontaneously or in response to 10⁻⁵ M ACh. Mean ± s . e . frequencies of spontaneous contractions ranged from 7·5 ± 2·0 h⁻¹ in the Atlantic silverside Menidia menidia to 380·3 ± 101·5 h⁻¹ in the sheepshead minnow Cyprinodon variegatus . Simultaneously tested ovaries from a single white perch Morone americana contracted at rates of 31·4 and 81·9 h⁻¹. This is consistent with independent generation of the contractile rhythm in the two ovaries. In three species (goldfish Carassius auratus , mummichog Fundulus heteroclitus and white perch), the tunica albuginea was removed from the ovary and found to be capable of independent contraction. This suggests that this membrane contributes to the observed contractions. The presence of ovarian contractions in this varied group of fish species indicates that the contractions play an important role in reproduction.     

Ploidy levels in silverside fishes (Atherinidae, Menidia) on the Texas coast: flow-cytometric analysis of the occurrence of allotriploidy

Flow cytometry of nuclear DNA fluorescence in erythrocytes, coupled with gene dosage assessments derived from electrophoresis of proteins, reaffirmed that the Menidia clarkhubbsi complex of unisexual atherinid fishes is diploid. Non-recombinant hybrids between M. beryllina and M. peninsulae represented 18% (108 specimens) of the Menidia collected from three pools in the Copano Bay area and 9-6% (42 specimens) of those from a pool on Galveston Island. Of those hybrids, 35 and 5%, respectively, were triploids. Zymograms indicated that the triploids either had two doses of genes from M. beryllina and one from M. peninsulas or they had one from the former species and two from the latter. The hybrids seem to be maintained primarily, if not entirely, by ongoing hybridization and back-crossing of F₁ hybrids to the parental species. Triploids apparently result from the combination of unreduced, diploid gametes from F₁ hybrids with haploid gametes from either M. beryllina or M. peninsulae.     

Scaling of oxygen consumption of Lake Magadi tilapia, a fish living at 37°C

Rates of oxygen consumption were measured in the geothermal, hot spring fish, Oreochromis alcalicus grahami by stopped flow respirometry. At 37° C, routine oxygen consumption followed the allometric relationship: V o₂=0.738 M ^0.75, where V o₂ is ml O₂ h ⁻¹ and M is body mass (g). This represents a routine metabolic rate for a 10 g fish at 37° C of 0.415 ml O₂ g⁻¹ h ⁻¹ (16.4 μmol O₂ g ⁻¹ h ⁻¹). Acutely increasing the temperature from 37 to 42° C significantly elevated the rate of O₂ consumption from 0.739 to 0.970 ml O₂ g ⁻¹ h ⁻¹ ( Q ₁₀=l.72). In the field, O. a. grahami was observed to be 'gulping' air from the surface of the water especially in hot springs that exceeded 40° C. O. a. grahami may utilize aerial respiration when O₂ requirements are high.     

Cytochrome c-mediated caspase-9 activation triggers apoptosis in Streptococcus pyogenes-infected epithelial cells

Epithelial cells are the initial sites of host invasion by group A Streptococcus pyogenes (GAS), and their infection of epithelial cells has been suggested to induce apoptosis. However, the mechanism responsible for bacteria–host interaction and the induction of apoptosis has not been clearly understood. We demonstrate here that human pharyngeal epithelial HEp-2 cells became apoptotic with DNA fragmentation by invasion of GAS strains JRS4 (M6⁺, F1⁺) and JRS145 (M6⁻, F1⁺ mutant of JRS4), whereas apoptotic cellular changes were not observed in SAM1 (M6⁺, F1⁻ mutant) or SAM2 (M6⁻, F1⁻ mutant) infected HEp-2 cells. Confocal microscopy revealed that Bax translocation to mitochondria and cytochrome c release occurred after 4 h of infection. Western blot analyses showed that the amounts of Bcl-2 and Bcl-x_L were decreased in the mitochondria of infected cells. In addition, we demonstrated that the release of nuclear histone from infected cells was prevented by the addition of caspase-9 inhibitor (Ac-LEHD-CHO). We conclude that the internalization of GAS in epithelial cells is necessary and sufficient for the induction of apoptosis, which is initiated by mitochondrial dysfunction, and the mechanism of GAS-induced apoptosis is clearly different from that induced by other intracellular invasive bacteria, e.g. Shigella and Salmonella species.     

The effect of dissolved oxygen and salinity on the toxicity of ammonia to smolts of salmon, Salmo salar L.

The survival of Atlantic salmon smolts on exposure to constant concentrations of ammonia has been measured under laboratory conditions. At concentrations of dissolved oxygen close to the air-saturation value, the 24-h LC50 of un-ionised ammonia is 0.15 mg NH₃1⁻¹ in fresh water (hardness 264 mg 1⁻¹ as CaCO₃) and 0.3 mg NH₃1⁻¹ in 30% sea water; at concentrations of dissolved oxygen of 3.5 mg 1⁻¹ in fresh water and 3.1 mg 1⁻¹ in 30% sea water, the 24-h LC50 is 0.09 mg NH₃ 1⁻¹ and 0.12 mg NH₃ 1⁻¹ respectively; for fish acclimated for 1 day to a concentration of ammonia close to the 24-h median for un-acclimated fish, the median is increased between 38 and 79%, depending on test conditions.     

Resistance to cadmium by pretreated rainbow trout alevins

A toxicity test with cadmium concentrations ranging from 0·1 to 100·0 mg Cd 1⁻¹ was used to assess the effect of cadmium pretreatment on rainbow trout ( Salmo gairdneri Richardson) alevins. The median period of survival for fish pretreated at 0·01 mg Cd 1⁻¹ was found to be increased at test concentrations up to 10mg Cd 1⁻¹ compared with alevins pretreated with dilution water. However, at concentrations above 10mg Cd 1⁻¹ pretreatment at 0·01 mg Cd 1⁻¹ reduced the median period of survival.     

Protoplasts from pectinolytic fungi: isolation, regeneration and pectinolytic enzyme production

S. Solís, M.E. FLORES AND C. HUITRON. 1996. Protoplast release in pectinolytic strain mutants of Aspergillus sp. CH-Y-1043 (A13) and Aspergillus flavipes ATCC-16795 (F7) is described. Optimum yield of protoplasts A13 was obtained in a lapse of 1 h when commercially lytic enzymes of Trichoderma harzanium (2 mg ml⁻¹) were added in 0.05 mol 1⁻¹ citrate-phosphate buffer pH 5.0 containing 0.7 mol 1⁻¹ KCl and 10 mg ml⁻¹ BSA. Best results in F7 were obtained when the protoplasting system of A13 was supplemented with 10 mg ml⁻¹ Aureobasidium sp. lytic enzymes. Isolated protoplasts in A13 and F7 were capable of a high regeneration frequency of 87% and 53% when 0.7 mol 1⁻¹ KCl and sorbitol were used as osmotic stabilizers. Endo-P, Exo-P and pectin lyase production were not modified during the process of regeneration.     

The rooting ability of shoots raised 'in vitro' from the apple rootstock A2 in juvenile and in adult growth phase

The apple rootstock A2 can be readily propagated in vitro both in the juvenile and in the adult growth phase. Shoots were produced by meristem tip culture from the apple rootstock A2 in different growth phases. The influence of growth phases and different concentrations of PG and IBA was investigated as to rooting percentage, survival percentage, number of roots per rooted shoot, root length, shoot length and formation of callus. IBA at 15 μ M without PG gave a significantly lower rooting percentage than 5 and 10 μ M IBA. PG together with IBA stimulated rooting, the optimum concentrations of PG being, however, not the same for the different growth phases. For the adult growth phase, 10⁻⁴ M PG promoted rooting, whereas 10⁻³ M PG markedly inhibited rooting. In the juvenile growth phases, both 10⁻⁴ and 10⁻³ M PG stimulated rooting. PG at 10⁻⁴ M also increased the number of roots. The longest roots were obtained at 10⁻³ M PG and 5 μ M IBA. PG at 10⁻³ M reduced callus formation at all IBA concentrations used. Neither shoot length nor root length influenced the survival percentage.     

Population ecology of pike-cichlid, Crenicichla lepidota, in two streams of the Brazilian Pampa subject to a severe drought

Scales were valid for age determination of pike-cichlid Crenicichla lepidota Heckel in two Brazilian streams, since, for every age-class, annual rings (annuli) were formed once a year, in August. Back-calculated Jengths-for-age coincided with those obtained by inspecting the monthly length-frequency distributions. Only 11 reproductive females were captured (length range 9.6–16 cm, age 1+ and 2 + ) and their fecundity ( F ) increased with fish length ( L ) according to: F= 2.4027 L^2.3631. Average annual density and biomass in four localities of Zanga do Campus were 2194, 1676, 1824 and 1071 individuals ha⁻¹ and 17.2, 12.1, 13.8 and 18.7 kg ha⁻¹and about 1000 individuals ha⁻¹ and 8.5 kg ha⁻¹ in three combined sites of Zanga do Barbará. The annual production ranged between 18.5 and 32.7 kg ha⁻¹ a ⁻¹ in the first stream and 8.7 kg ha ⁻¹a ⁻¹ in the other. Feeding tactics differed between streams and age-classes (0+ v . 1+/2+). In winter, the 0+ age-class in Zanga do Campus fed mainly upon Ephemeroptera (Baetidae) but in other months also fed on zooplankton. In winter, these age-classes in Zanga do Barbara preyed upon benthic species but the contribution of different taxa varied between months. In January, these fish preyed exclusively upon Chydoridae and fish. The effects of a severe drought (November-February) upon these parameters are discussed.     

Olfactory sensitivity of the herbivorous grass carp, Ctenopharyngodon idella, to amino acids

The olfactory sensitivity of a herbivore, the grass carp, Ctenopharyngodon idella , to ten amino acids was investigated using electrophysiological recording techniques. Underwater electroolfactogram recordings increased exponentially with logarithmic increases of stimulus concentration from threshold to 1 × 10⁻³ m. Calculated detection thresholds ranged from 1 × 10^−7.15 m to 1 × 10^−8.7 M and are similar to those previously reported for carnivorous fish species. The relative stimulatory efficacy (RSE) of the ten amino acids determined at 1 × 10⁻⁴ M revealed five groups of stimuli, with cysteine the most stimulatory and proline the least. Measures of amino acid RSE did not differ significantly from those of previously studied carnivores, except for arginine. Grass carp showed strong responses to this compound, whereas it has produced only limited responses in carnivores. Differences in the responsiveness of herbivores and carnivores were not observed, suggesting that feeding niche segregation is probably not facilitated by differential olfactory sensitivities to feeding stimuli.     

Effects of phenolic acids on ammonia oxidation by terrestrial autotrophic nitrifying microorganisms

Abstract It has been hypothesized that vegetation in certain ecosystems inhibits nitrification in soil by producing phenolic compounds that inhibit oxidation of ammonia by nitrifying microorganisms. This hypothesis is based largely on a report that very low concentrations (10⁻⁶ M–10⁻⁸ M) of several phenolic acids (notably ferulic acid) completely inhibited NO₂⁻ production in an aqueous suspension of soil treated with (NH₄)₂SO₄ and a nutrient solution suitable for growth of Nitrosomonas and other autotrophic nitrifying microorganisms. To evaluate this hypothesis, we determined the effects of three ohenolic acids (ferulic acid, caffeic acid, and p -coumaric on nitrite production by representatives of three genera of terrestrial autotrophic nitrifying microorganisms ( Nitrosospira, Nitrosomonas , or Nitrosolubos ) grown on a defined medium containing NH₄⁺. We found that nitrite production by the Nitrososspira was not inhibited by ferulic acid, caffeic acid, or p -coumaric acid at concentrations of 10⁻⁶ or 10⁻⁵ M and was only slightly inhibited when these acids were at a concentration of 10⁻⁴ M. We also found that ferulic acid did not markedly inhibit nitrite production by the three genera of nitrifying microorganisms studied, even when its concentration was as high as 10⁻³ M. These observations invalidate the hypothesis tested because the phenolic acids studied did not significantly retard ammonia oxidation by autotrophic microorganisms even when their concentration in cultures of these microorganisms greatly exceeded their concentrations in soils.     

Photosynthesis and photoassimilation of glucose during photomixotrophic growth of Marchantia polymorpha cells in suspension culture

Even in the presence of glucose the growth of Marchantia polymorpha L. (cell line HYH-2F) requires light, and growth is more sensitive to 10⁻⁶ M 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea than to 10⁻⁴ Antimycin A. The inability of the cells to grow in the dark is due to the low level of respiration. The respiration rate under light increased to four times the dark value. The values of the compensation ratio (the photosyntehtic rate/the respiration rate) for the oxygen exchange were below 1.0 daring the growth period, although oxygen evolution was found. At the early exponential phase, oxygen evolution was 0.373 μmol (mg cell dry weight)⁻¹ h⁻¹ [61.7 μmol (mg chlorophyll)⁻¹ h⁻¹]. M. polymorpha cells are unable to grow anaerobically in the light without a supply of carbon dioxide. When 1% carbon dioxide in nitrogen is supplied, photochemically produced oxygen and energy are sufficient for sustained growth although at significantly reduced yields in both cell dry weight and chlorophyll. Photosyntehtic CO₂ assimilation rate was 0.13 μmol (mg cell dry weight)⁻¹ h⁻¹[11.3 μmol (mg chlorophyll)⁻¹ h⁻¹]. At least one-third of the carbon atoms in cellular constituents seem to be derived from atmospheric carbon dioxide, which indicates that M. polymorpha cells grow photomixotrophicaily.     

Some effects on the growth of brown trout from exposure to aluminium at different pH levels

Yearling brown trout, Salmo trutta L., were exposed to various concentrations of inorganic aluminium (0–3.7 μM1⁻¹) over a pH range of 4.3–6.5 in a flow-through bioassay apparatus using synthetic test media. Low pH, in the absence of aluminium, produced little effect on growth or survival except at the lowest pH tested (4.3). At pH less than 5.5, concentrations of total aluminium in excess of 1 μM 1⁻¹ (27μg 1⁻¹) were found to retard growth. The effects of a given aluminium concentration were markedly reduced at pH above 5.5.
The change in aluminium toxicity with pH must be related to changes in aluminium chemistry. When growth rates are correlated with the different aluminium species, calculated using thermodynamic equilibrium constants given in the literature, it appears that the Al(OH)^{2 +} species is the most toxic, with a small contribution also coming from polymeric complexes.     

Attraction of zebrafish, Brachydanio rerio, to alanine and its suppression by copper

Preference responses of zebrafish to 10⁻³, 10⁻⁴ and 10⁻⁵M alanine (Ala) were concentration- dependent. Behavioural responses to copper (Cu) and Cu + Ala mixtures were also assessed. Zebrafish avoided 100 and 10 μg Cu l⁻¹, but not 1 μg l⁻¹. Mixtures of 10⁻³ m Ala+ 100 μg Cu l⁻¹ and 10 ⁴ M Ala + 10 μg Cu 1⁻¹ were avoided as intensely as was Cu alone. Responses to 10⁻³ M Ala + 10 or 1 μg Cu l⁻¹ and 10 ⁴ M Ala +1 μg Cu l⁻¹ did not differ statistically from controls (no detectable preference or avoidance). These results demonstrate, firstly, that a concentration of a pollutant avoided by itself (10 μg Cu l⁻¹) may not be avoided when encountered with an attractant chemical stimulus (Ala) and may suppress the preference for an attractant stimulus, and secondly, that a concentration of a pollutant not avoided by itself and not considered deleterious (1 μg Cu l⁻¹) suppresses attraction to Ala (an important constituent of prey odours for many fishes).     

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.     

The relationship of diet to growth and ammonium excretion in salt marsh fish

Synopsis Detritus is an abundant but poor quality food source for consumers in salt marsh ecosystems. Here we present results of feeding experiments to determine the ability of Fundulus heteroclitus, Cyprinodon variegatus, and Mugil cephalus, three major detritivores in Great Sippewissett Marsh, Massachussetts, to assimilate detritus and use it for growth. C. variegatus, the sheepshead minnow, gained weight on a detrital diet, but F. heteroclitus, the marsh killifish, and juvenile M. cephalus, the striped mullet, lost weight and suffered high mortality on detrital diets. C. variegatus is a herbivore with morphological adaptations for ingesting plant material. F. heteroclitus is a carnivore poorly suited to effectively assimilate detritus from the diet. Although adult M. cephalus are adapted for ingesting and assimilating detrital material, the young may lack these adaptations and thus do not assimilate detritus. C. variegatus excretes ammonium at a lower rate than F. heteroclitus when fed animal food, or when starved for short periods of time. This protein sparing effect could be crucial for survival when detritus is the only food available at certain times of the year. While both C. variegatus and M. cephalus are visitors in the marsh and may leave to exploit food sources elsewhere, F. heteroclitus is a year-round resident and is the most abundant species of fish in the marsh. Yet when high quality food sources become scarce in late summer and fall, detritus, although plentiful, is apparently not a suitable alternative.To whom all correspondences should be addressed.     

Amount and distribution of biochemical-genetic variation among wild populations and a hatchery stock of Atlantic salmon, Salmo salar L., from north-east Ireland

The effects of tributyltin (TBT) compounds on gill morphology were examined in the mummichog, Fundulus heteroclitus , in 96-h LC50 and 6-week sublethal exposures. Morphometry was used for the identification and quantification of effects with the light microscope. A 96-h LC50 of 17.2 μg 1 ¹ was determined. Morphometric analysis of gill tissues revealed hypertrophy of the lamellar epithelium in fish exposed to 17.2 μg 1–1. Relative diffusing capacity was significantly decreased (−41 %); ( P <0.05, ANOVA, Bonferroni t -test). At 35.6 μg 1⁻¹, TBT exposure resulted in a significant reduction (− 40%) in the volume of the lamellar blood channels. Both of these observations occurred in fish that showed signs of acute poisoning including loss of equilibrium. In fish exposed to sublethal concentrations of 0.105–2.000 μg TBT 1⁻¹ for 6 weeks, there were no pathological changes in the gill. There were no treatment-related changes in the surface morphology of the gills of fish from both experiments upon scanning electron microscopic examination. Although gill pathology was observed in acutely toxic exposures, it does not appear to be a major mechanism of TBT toxicity.     

Identification of jasmonic acid in Mimosa pudica and its inhibitory effect on auxin- and light-induced opening of the pulvinules

Jasmonic acid was identified from Mimosa pudica L. plants by mass spectrometry, high performance liquid chromatography and thin layer chromatography. Effects of authentic jasmonic acid on pulvinule movement and transpiration of the pinnae were compared with those of abscisic acid. Jasmonic acid and abscisic acid each at 10⁻⁵ M inhibited both auxin- and light-induced opening of the pulvinules. A closure-inducing activity of jasmonic acid at 10⁻⁴ M was greater than that of abscisic acid at 10⁻⁴ M. Pinnae transpiration was reduced by 10⁻⁵ M abscisic acid but not by 10⁻⁴ M jasmonic acid.     

In vitro carbon dioxide excretion from erythrocytes of two species of Antarctic fishes and its inhibition by catecholamines

This study was designed to investigate whether the blood of Pagothenia borchgrevinki , exhibits a Haldane effect, and whether activation of a Na⁺/H⁺ antiporter increases transport of intracellular protons and Bohr protons out of the erythrocytes resulting in inhibition of CO₂ excretion in both P. borchgrevinki , and Dissostichus mawsoni. When carbon dioxide dissociation curves were determined from blood samples pooled from three fish under oxygenated and deoxygenated conditions a Haldane effect was observed. Using an in vitro , CO₂ excretion assay, the rate of HCO₃⁻ dehydration was determined on blood and plasma equilibrated under an N₂atmosphere then rapidly oxygenated with air in the presence of 10⁻⁵ M noradrenaline or acetazolamide (10⁰⁴M). Whole blood and plasma from P. borchgrevinki , and D. mawsoni , were equilibrated with 0·5% CO₂ in air and assayed in the presence of 10⁻⁵ M noradrenaline. Erythrocyte CO₂ excretion rates were depressed significantly by noradrenaline in both species. The whole blood HCO₃⁻ dehydration rate was depressed significantly following rapid oxygenation in the presence of acetazolamide indicating that the pathway of CO₂ excretion included activation of intracellular carbonic anhydrase and an adrenergic receptor.     

Taste responses of the facial and glossopharyngeal nerves to amino acids in the rainbow trout

No significant differences were noted between responses of rainbow trout Oncorhynchus mykiss facial and glossopharyngeal nerves to 15 amino acids. Nine of these amino acids tested at 10⁻² M were stimulatory, whereas only two tested at 10⁻³ M were effective gustatory stimuli. For both nerve systems, ≤10⁻³ M L-proline was the most stimulatory amino acid, with an estimated threshold of 10⁻⁷ M; however, L-α-amino-β-guanidino-propionic acid (estimated threshold of 3×10⁻³ M), was the most potent compound at 10⁻² M. These results indicate that the same amino acids activate taste buds innervated by facial and glossopharyngeal nerves, respectively, and suggest that the same amino acids can be important in chemosensory feeding behaviour in the rainbow trout.