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).     

Entrainment to light of circadian activity rhythms in tench (Tinca tinca)

The present article analyzes locomotor activity rhythms in Tinca tinca. To that end, three different experiments were conducted on 24 animals (20 g body weight) kept in pairs in 60-liter aquaria fitted with infrared sensors connected to a computer to continuously record fish movements. The first experiment was designed to study the endogenous circadian clock under free-running conditions [ultradian 40:40 min LD pulses and constant dark (DD)] and after shifting the LD cycle. Our results demonstrate that tench has a strictly nocturnal activity pattern, an endogenous rhythm being evident in 45.8% of the fish analyzed. The second experiment was conducted to test the influence of different photoperiods (LD 6:18, 12:12, 18:6, and 22:2) on locomotor activity, the results showing that even under an extremely long photoperiod, tench activity is restricted to dark hours. The third experiment examined the effect of light intensity on locomotor activity rhythms. When fish were exposed to decreasing light intensities (from 300:0 lux to 30:0, 3:0, and 0.3:0 lux) while maintaining a constant photoperiod (LD 12:12), the highest percentage of locomotor activity was in all cases associated with the hours of complete darkness (0 lux). In short, our results clearly show that (a) tench is a species with a strictly nocturnal behavior, and (b) daily activity rhythms gradually entrain after shifting the LD cycle and persist under free-running conditions, pointing to their circadian nature. However, light strongly influences activity rhythms, since (c) the length of the active phase is directly controlled by the photophase, and (d) strictly nocturnal behavior persists even under very dim light conditions (0.3 lux). The above findings deepen our knowledge of tench behavior, which may help to optimize the aquacultural management of this species, for example, by adjusting feeding strategies to their nocturnal behavior.     

The effects of tench (Tinca tinca (L.)) and sticklebacks (Gasterosteus aculeatus L.) on planktonic and benthic communities in mesocosms in a shallow lake

The effects of introducing a zooplanktivorous fish, three-spined stickleback, (Gasterosteus aculeatus) and a benthivorous fish, tench (Tinca tinca) separately and in combination to replicated experimental enclosures with two density levels of white water lily (Nymphaea alba) were studied in Little Mere, UK. Numbers of Daphnia hyalina were high and only slightly diminished at reduced lily densities, probably due to stickleback predation, but there was no consequential effect on phytoplanktonic chlorophyll a concentrations. Tench reduced the numbers of gastropods but not of other macroinvertebrates, and in turn increased the biomass of periphyton growing on artificial substrata within the enclosures. The higher lily density reduced oxygen concentrations and pH values and increased total phosphorus and soluble reactive phosphorus concentrations but otherwise had little effect on water chemistry. There was little interactive effect of the fish species. The results are integrated with those of six other such enclosure experiments carried out in Little Mere since 1992.     

Seasonal changes in phagocytic capacity and superoxide anion production of blood phagocytes from tench (Tinca tinca,L.)

Seasonal variations in the ex vivo phagocytic function of blood cells from tench, including ingestion capacity of inert particles and its destruction (microbicide capacity) assessed by measurement of superoxide anion production, were studied. Tench were maintained under natural conditions throughout the year, and the different assays of samples taken during each season were initially performed in vitro at 22°C and the results compared. Subsequently, assays were performed at the same temperature as that of the water ponds in which the fish were kept (“seasonal temperature”: 12°C in winter, 22°C in spring and autumn and 30°C in summer) and the results compared seasonally. The results at 22°C showed that phagocytic capacity was greatest in spring and summer and lowest in winter. However, when phagocytic capacity was measured at seasonal temperature, highest values appeared in winter and lowest in summer and autumn. Nitroblue tetrazolium reduction by tench phagocytes after phagocytosing latex beads demonstrated a similar seasonal behaviour at both 22°C in each season and at seasonal temperature. The highest values appeared in summer, which suggests a better microbicide capacity in this season. The results obtained in this study suggest that for a correct interpretation of ex vivo phagocytic capacity of fish through the year it is necessary to use the same assay temperature as that of the water in which the fish is kept.     

Diet composition and prey preference of tench, Tinca tinca (L.), common bream, Abramis brama (L.), perch, Perca fluviatilis L. and roach, Rutilus rutilus (L.), in two contrasting gravel pit lakes: potential trophic overlap with wildfowl

The abundance of benthic macroinvertebrate taxa and the relative abundance of zooplankton taxa were compared with the diet of bream, tench. perch and roach from two gravel pit lakes during May-July 1986 and 1987. Significant food preferences were demonstrated between species and between lakes. Chironomid pupae dominated the macroinvertebrate diet of perch, roach, ate predominantly either Spirogyra sp. (St Peters Lake) or Daphniu hyalina (Main Lake).
The Main Lake, bream fed largely upon chironomid larvae and the bivalve Sphuerium but in St Peters Lake they positively selected a variety of less abundant benthic invertebrate taxa. Bream switched from benthos to zooplankton in the Main Lake in 1986. Tench ate large numbers of Aselhs and showed positive selection of various macroinvertebrate prey in St Peters but ate D. hjulim in Main Lake. Tropic overlap for chironomids and other macroinvertebrate prey was demonstrated between perch, bream and tench, and potentially with wildfowl which used the gravel pits for breeding and wintering.     

Entrainment to Light of Circadian Activity Rhythms in Tench (Tinca tinca)

The present article analyzes locomotor activity rhythms in Tinca tinca. To that end, three different experiments were conducted on 24 animals (20 g body weight) kept in pairs in 60‐liter aquaria fitted with infrared sensors connected to a computer to continuously record fish movements. The first experiment was designed to study the endogenous circadian clock under free‐running conditions [ultradian 40:40 min LD pulses and constant dark (DD)] and after shifting the LD cycle. Our results demonstrate that tench has a strictly nocturnal activity pattern, an endogenous rhythm being evident in 45.8% of the fish analyzed. The second experiment was conducted to test the influence of different photoperiods (LD 6:18, 12:12, 18:6, and 22:2) on locomotor activity, the results showing that even under an extremely long photoperiod, tench activity is restricted to dark hours. The third experiment examined the effect of light intensity on locomotor activity rhythms. When fish were exposed to decreasing light intensities (from 300:0 lux to 30:0, 3:0, and 0.3:0 lux) while maintaining a constant photoperiod (LD 12:12), the highest percentage of locomotor activity was in all cases associated with the hours of complete darkness (0 lux). In short, our results clearly show that (a) tench is a species with a strictly nocturnal behavior, and (b) daily activity rhythms gradually entrain after shifting the LD cycle and persist under free‐running conditions, pointing to their circadian nature. However, light strongly influences activity rhythms, since (c) the length of the active phase is directly controlled by the photophase, and (d) strictly nocturnal behavior persists even under very dim light conditions (0.3 lux). The above findings deepen our knowledge of tench behavior, which may help to optimize the aquacultural management of this species, for example, by adjusting feeding strategies to their nocturnal behavior.     

Determination of plasma melatonin levels by enzyme-linked immunosorbent assay (EIA) in turbot (Scophtalmus maximus L.) and tench (Tinca tinca L.)

An enzymoimmunoassay (EIA) kit for plasma melatonin (MLT) measurements was employed in tench (Tinca tinca) and in turbot (Scophtalmus maximus). Tench and turbot plasma samples were purified with a C18 reversed phase extraction columns because this kit is designed for human serum measurements. The lowest detection limit of the technique was 11.48 pg/well with a sensitivity at 50% binding of 100 pg/well. Intra-assay and inter-assay CV (%) were always less than 5% (n=8), and 9% (n=6) in tench plasma samples, and less than 5% (n=8) and 13% (n=5) in turbot plasma samples, respectively. Correlation coefficients between EIA and RIA measurements in tench and turbot plasma samples were 0.93 and 0.89 (p<0.001) respectively. Diurnal and nocturnal plasma melatonin mean levels were 14.7+/-2.1 pg/ml and 87.4+/-11 pg/ml in tench (n=15), and 3.5+/-0.4 pg/ml and 28.1+/-2.1 pg/ml in turbot (n=15). These species showed a melatonin circadian rhythm as in other animals studied. The results suggest that the commercial kit used in this experiment could be a suitable and alternative method to RIA for plasma MLT determinations in tench and turbot although it is necessary to increase volumes (1ml) and concentrate daytime samples.     

Egg incubation time and hatching success in tench Tinca tinca (L.) related to the procedure of egg stickiness elimination

The experiment showed different results after a short (2 min) enzyme alcalase Merck EC 3.4.21.14 (5.0 ml L^?1 concentration) treatment of tench eggs in contrast to the traditional methods of eliminating egg stickiness involving milk solution (50 g L^?1) treatment for 70 min followed by the addition of a talc suspension (33 g L^?1) for 10 min or treatment by fine clay suspension (20 g L^?1) for 60 min or talc suspension (33 g L^?1) for 80 min. The alcalase enzyme treatment resulted in decreased egg stickiness compared with the conventional milk/clay/talc treatments, indicated by lower duration of egg incubation and higher hatching rates (anova for hatching rate, P < 0.0084). The highest hatching rate (93.2%) was achieved using the enzyme; the lowest (31.3%) was using a talc suspension (control hatching rate was 86.2%). Duration of egg incubation at degree‐days (D°) after enzyme treatment (58.6 D°) was about 4–5 h shorter than the classical method using milk solution and talc suspension (63–65 D°). Prolongation in the latter classical method may also be explained by a hardening of the egg envelopes.     

Survival and changes in growth of juvenile tench (Tinca tinca L.) fed defined diets commonly used to culture non-cyprinid species

The study is an attempt to evaluate the feasibility of intensive tench culture using non‐specific diets as a preliminary step to check the acclimatization of the species under intensive rearing systems. Five‐month‐old juvenile tench were reared in recirculating systems at mean water temperatures of 22°C for 75 days. Fish fed with four different commercial diets (trout starter, trout first feeding, sea‐bass and eel), showed significantly higher final weights than the fish fed either eel or sea‐bass diets. Initial weight for all treatments was 2.3 ± 0.53 g. Final weight for the commercial diet groups was 3.56 ± 0.4 g, compared with the remaining groups that reached 2.09 ± 0.47 g (P < 0.05). Significantly higher survival rates were observed in the eel and sea‐bass groups (84.7 and 51.5%, respectively) than in either of the trout diet groups (38%). Specific growth rates (1.26 vs –0.18) and condition factor (1.26 vs 0.93) were also higher than those fed with salmonid diets (P < 0.05). Results obtained in this study indicate that regardless of the species’ slow growth, when compared with other cyprinids, final growth rates and survival of tench fed exclusively on sea‐bass or eel diets can be considered satisfactory. It must be pointed out that these promising results were obtained at lower temperatures than previous studies of tench in culture systems. The use of belt feeders did not show improvement in growth compared with manually fed fish when trout diets were used.     

Buoyancy variations in eggs of Atlantic cod (Gadus morhua L.) in relation to chorion thickness and egg size: theory and observations

Egg specific gravity is a function of egg volume, chorion volume, perivitelline space (PVS) and the specific gravity of chorion and ovoplasm. Experimental studies on cod ( Gadus morhua L.), a marine multiple batch spawner, showed that approx. forty per cent of the chorion volume is incorporated between batches and that the chorionic material has a mean specific gravity of 1·20 (range: 1·14–1·35). The chorion volume and specific gravity are low at the end of the spawning period in extremely fecund fish. Specific gravity of ovoplasm was observed to be 1·017 in marine eggs and calculated to be about 1·008 in brackish water eggs. Inherent variation in PVS is found to have little influence on egg specific gravity. With the other variables constant, egg specific gravity is inversely related to egg volume supported by studies on single batches of eggs. Measurements on every batch of eggs reveal that the production of large eggs (>1·3 mm) gives low density eggs initially and high density eggs towards the end of spawning, while the production of small eggs (<1·3mm) gives moderate specific gravity. These observations are discussed in relation to vertical distribution of the eggs and reproductive success of recruit and repeat spawners. There are indications that repeat spawners produce larger eggs.     

Effect of antibody to detergent solubilized zonae pellucidae on in vivo and in vitro fertilization in the mouse

An antiserum was produced in one rabbit against mouse zonae pellucidae solubilized with 70 mM Na₂SO₃, 1% SDS, and 0.04 mM CuSO₄. An IgG of zona antibody completely inhibited fertilization both in vitro and in vivo in the mouse. F(ab′)₂ fragment obtained by pepsin digestion of IgG zona antibody inhibited fertilizability of eggs in vitro but did not inhibit fertilization in vivo after passive immunization.     

In vivo and in vitro flowering response of chicory (Cichorium intybus L.): influence of plant age and vernalization

Chicory plants (Cichorium intybus L. var foliosum cv Flash) were tested with and without a 4-week-long cold treatment for in vivo and in vitro flowering potential every 2 weeks during the growing season. One hundred percent of the plants harvested 112 days or later after sowing and then vernalized flowered in vivo. In vitro, no vernalization was needed to initiate flowering-stems on chicory explants taken from roots of 100 days old and older. 5-Azacytidine, a DNA demethylation agent, increased the flowering percentage on explants from young, vernalized roots but could not induce more than 15% flowering on young, nonvernalized roots. The greater flowering potential of chicory root explants in vitro when compared to plants of the same age tested in vivo was clearly established. This result suggests that some negative control on flowering was removed when root explants were excised and the main plant body discarded. Received: 31 August 1998 / Revision received: 27 October 1998 / Accepted: 10 November 1998     

Reproductive division of labour and thelytoky result in sympatric barriers to gene flow in honeybees (Apis mellifera L.)

Determining the extent and causes of barriers to gene flow is essential for understanding sympatric speciation, but the practical difficulties of quantifying reproductive isolation remain an obstacle to analysing this process. Social parasites are common in eusocial insects and tend to be close phylogenetic relatives of their hosts (= Emery's rule). Sympatric speciation caused by reproductive isolation between host and parasite is a possible evolutionary pathway. Socially parasitic workers of the Cape honeybee, Apis mellifera capensis, produce female clonal offspring parthenogenetically and invade colonies of the neighbouring subspecies A. m. scutellata. In the host colony, socially parasitic workers can become pseudoqueens, an intermediate caste with queenlike pheromone secretion. Here, we show that over an area of approximately 275.000 km2, all parasitic workers bear the genetic signature of a clone founded by a single ancestral worker genotype. Any gene flow from the host to the parasite is impossible because honeybee workers cannot mate. Gene flow from the parasite to the host is possible, as parasitic larvae can develop into queens. However, we show that despite sympatric coexistence for more than a decade, gene flow between host and social parasite (F(st) = 0.32) and hybridizations (0.71%) are rare, resulting in reproductive isolation. Our data suggest a new barrier to gene flow in sympatry, which is not based on assortative matings but on thelytoky and reproductive division of labour in eusocial insects, thereby suggesting a new potential pathway to Emery's rule.     

Suppressive effect of ethanol extract from mango (Mangifera indica L.) peel on IgE production in vitro and in vivo

Abstract

Immunoglobulin E (IgE) is involved in the onset of allergic reaction, and the suppression of IgE production leads to alleviation of allergic symptoms. We found that mango peel ethanol extract (MPE) significantly suppresses IgE production by human myeloma cell line U266 cells, suggesting that MPE has an anti-allergic effect by inhibiting the production of IgE. Although mangiferin is contained in mango, which suppresses IgE production by U266 cells, it was not contained in MPE. We investigated the suppressive effect of MPE in 2,4-dinitrofluorobenzene (DNFB)-induced allergic contact dermatitis model mice. The elevation of serum IgE level was significantly suppressed by oral administration of MPE. Intake of MPE also suppressed the expression level of IL-4 in the DNFB-challenged ears, suggesting that MPE suppresses the IL-4-mediated maturation into IgE-producing cells. Our findings indicate that MPE has a potential to alleviate the increase in serum IgE level that is feature of type I allergy.     

Genotypic influence on in vitro induction, dormancy length, advancing age and agronomical performance of potato microtubers (Solanum tuberosum L.)

Microtubers of 13 cultivars, largely grown in Italy and other European countries, were induced. They were stored in the dark at 3°C for different periods (28, 56, 84 and 105 days), prior to being transferred to 20°C for between 4 and 17 weeks. Following removal to room temperature, sprouting was recorded and dormancy duration quantified. Dormancy decreased from 28.1 to 19.9, 11.1 and 7.8 days with reduced time of storage. Cvs Arsy, Nicola and Jaerla took consistently more time for dormancy release. The dormancy duration was linearly and inversely correlated with the length of storage. After sprouting, tubers were held at 20°C for various intervals and a range of physiological ages (0, 368, 720 and 1008 degree days) were accumulated. The field comparison of microtubers evidenced a plant growth response and tuber yield/plant affected by the cultivar and physiological age. In early cultivars (Jaerla), a better performance was shown by younger tubers; the opposite trend was noted in Alpha (a later cultivar) with an increase in stems/plant, tubers/plant and tuber yield/plant for tubers with greater physiological age. Like conventional seed tubers, microtubers showed differences in optimum physiological age associated with cultivar earliness. This study has provided some indications on how to enhance emergence and haulm development of plants from microtubers.     

Antifungal activity of the dill (Anethum graveolens L.) seed essential oil against strawberry anthracnose under in vitro and in vivo conditions

Strawberry anthracnose, caused by Colletotrichum nymphaeae, is mainly controlled by the application of synthetic chemical fungicides. The present study assessed the antimicrobial activity of essential oils (EOs) from dill (Anethum graveolens L.) seed against C. nymphaeae. The antifungal effects of dill seed EO on C. nymphaeae was initially evaluated in vitro and further extended as in vivo condition. The results indicate that in the contact and volatile assays, dill seed EO significantly inhibited mycelial growth of C. nymphaeae at all concentrations tested. Conidia germination was also significantly inhibited at concentrations of 250 – 1000 ppm. Disease incidence and severity of anthracnose on strawberry fruits were significantly reduced compared with infected control, from concentrations of 500 and 50 ppm, respectively. The results confirm the efficacy of dill seed EO against C. nymphaeae, which may represent an alternative to synthetic chemical fungicides to control strawberry anthracnose pre- and post-harvest.     

In vitro and in vivo screening for the identification of salt-tolerant sugarcane (Saccharum officinarum L.) clones: molecular,biochemical, and physiological responses to salt stress

Sugarcane is a glycophyte whose growth and yield can be negatively affected by salt stress. As the arable lands with potential saline soils expand annually, the increase of salt-tolerance in sugarcane cultivars is highly desired. We, herein, employed in vitro and in vivo conditions in order to screen sugarcane plants for salt tolerance at the cellular and at the whole plant levels. Calli of sugarcane cv. Khon Kaen 3 (KK3) were selected after culturing in selective media containing various NaCl concentrations, and regenerated plants were then reselected after culturing in selective media containing higher NaCl concentrations. The surviving plants were finally selected after an exposure to 254 mM NaCl under greenhouse conditions. A total of 11 sugarcane plants survived the selection process. Four plants that exhibited tolerance to the four different salt concentrations applied during the aforementioned screening process were then selected for the undertaking of further molecular, biochemical, and physiological studies. The construction of a dendrogram has revealed that the most salt-tolerant plant was characterized by the lowest genetic similarity to the original cultivar. The relative expression levels of six genes (i.e., SoDREB, SoNHX1, SoSOS1, SoHKT, SoBADH, and SoMIPS) were found to be significantly higher in the salt-tolerance clones than those measured in the original plant. The measured proline levels, the glycine betaine content, the relative water content, the SPAD unit, the contents of chlorophyll a and b, as well as the K⁺/Na⁺ ratios of the salt-tolerant clones were also found to be significantly higher than those of the original plant.When the salt-tolerant clones were grown in a low saline soil, they exhibited a higher Brix percentage than that of the original cultivar.     

Effect of fertilization on exudation,dehydrogenase activity,iron-reducing populations and Fe++ formation in the rhizosphere of rice (Oryza sativa L.) in relation to iron toxicity

Summary To explain the mechanism of iron toxicity, greenhouse and growth chamber (¹⁴CO₂ atmosphere) experiments were carried out. In pot experiments (with a typical iron-toxic soil and a fertile clay) we studied the effect of N, P, K and Ca+Mg fertilization (alone or in combination) on dehydrogenase activity, Fe⁺⁺ formation, and the populations of iron-reducing bacteria in the rhizosphere of rice IR22 and IR42. Fe uptake by the plants was measured at regular intervals. Dehydrogenase activity, the number of N₂-fixing iron-reducing bacteria, and the formation and uptake of Fe⁺⁺ decreased with increased supply of K, Ca, and Mg. This effect was clearer with IR22 (susceptible to iron toxicity) than with IR42 (releatively tolerant). Increased exudation and Fe uptake by IR36 at low nutrient and high Fe supply were recorded in a growth chamber experiment. Nutritional conditions, exudation rate (a measure of metabolic root leakage), the iron-reducing activity of the rhizosphere, and Fe⁺⁺ uptake by wetland rice appear to be clearly related. Iron toxicity is considered a physiological disorder caused by multiple nutritional soil stress rather than by a low pH and high Fe supply per sé.     

Characterisation of the storage protein subunits synthesised in vitro by polyribosomes and RNA from developing pea (Pisum sativum L.)

Evidence is presented to show that legumin, the major storage protein in Pisum, is synthesised in vitro by the wheat germ and reticulocyte lysate systems, from polyribosomes and mRNA isolated from developing pea seeds. While legumin isolated from mature pea seeds consists of 40,000 and 20,000 MW subunits, the in vitro legumin is synthesised as a 60,000 MW precursor consisting of covalently linked 40,000 and 20,000 MW subunits. The implications of these findings are discussed in relationship to studies with other systems.Abbreviations SDS-PAGE SDS-polyacrylamide gel electrophoresis - PBS phosphate buffered saline - MW molecular weight