Changes in the selected hematological parameters and gill Na/K ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery

The increase in concentration of ammonia in lake water during the degradation of algal blooms may last for several weeks and thus cause chronic toxicity to aquatic organisms. The purpose of this study was to assess the chronic toxicity of ammonia on the selected hematological parameters and gill Na⁺/K⁺ ATPase activity of juvenile crucian carp Carassius auratus during elevated ammonia exposure and the post-exposure recovery. Juvenile crucian carp were exposed in different ammonia solutions for 45 days and then immediately transferred to pristine freshwater to initiate a 15-day recovery period. Results showed sub-lethal ammonia significantly deters growth and a 15-day recovery period was not sufficient for the fish to compensate for the loss of growth. The fish exhibited a continuous decrease in red blood cell (RBC), the total hemoglobin (Hb), and gill Na⁺/K⁺ ATPase activity as the concentration of NH₃-N increased. After the 15-day recovery period, RBC, Hb, and gill Na⁺/K⁺ ATPase activity had recovered to similar levels as the controls.     

Ontogeny and disease responses of Langerhans-like cells in lymphoid tissues of salmonid fish

The ontogeny and disease responses of Langerhans-like cells within lymphoid tissues of Atlantic salmon, Salmo salar, and rainbow trout, Oncorhynchus mykiss, were investigated. These cells were studied in situ with the use of two markers: the ultrastructural presence of Birbeck-like granules and immunohistochemistry with an antibody against human langerin/CD207 that cross-reacts with salmonid tissues. The appearance of Birbeck-like granules was observed in rainbow trout at 2 weeks post-hatch (PH) in the thymus and anterior kidney prior to the development of the spleen. Spleen first appeared at 3 weeks PH in both Atlantic salmon and rainbow trout, and Birbeck-like granules were observed within cells of the newly developed spleens. The cross-reactivity of langerin as seen by immunohistochemistry was not clearly observed in kidney and spleen until 9 weeks PH, when a strong cytoplasmic reaction was observed. To study langerin-positive cells in spleen and kidney during disease, microsporidial gill disease (MGD) in rainbow trout was used as a known disease model inducing a strong cell-mediated adaptive immune response. Langerin-positive cells in healthy fish were seen predominantly in the spleen, and only low numbers were present in the anterior kidney. During MGD, langerin-positive cell numbers were elevated in the anterior kidney and were significantly higher during 5, 6, and 10 weeks post-exposure (PE) compared with healthy control tissue. During MGD, the distribution of langerin-positive cells in the spleen and anterior kidney shifted from having significantly higher numbers of cells in the spleen than in the kidney in controls and at 1 and 4 weeks PE to having a similar distribution of the cells in the two organs at 2, 3, 5, and 6 weeks PE. By 10 weeks PE, significantly higher numbers of langerin-positive cells occurred in the anterior kidney compared with the spleen.     

Responses of the killifish (Aphanius dispar) to long-term exposure to elevated temperatures: growth,survival and microstructure of gill and heart tissues

Some fish species, such as killifish, that normally inhabit temperate water environments are also found in extreme thermal environments such as thermal springs. The extent of the adaptations involved is not known. In the present laboratory study, we exposed killifish (Aphanius dispar) acclimated to a normal thermal environment to elevated temperatures (37–40 °C) in which related killifish species live permanently. Our objective was to determine whether there is evidence that killifish have heat-shock characteristics that make permanent adaptation likely. The fish was exposed to this temperature for a period of 44 days and then compared with control fish kept at their normal temperature (24 °C) with respect to growth, survival and histopathology of gill and heart tissues. At the end of the experimental time, the percentage of body weight gain and specific growth rate were significantly lower in fish kept in thermal stress compared with the control group. Feed conversion ratio (FCR) was also significantly affected by water temperature, so that during thermal stress the values of FCR were negative. Fish condition (Condition factor: CF) did not differ significantly between both groups at the end of the experiment. On days 11 and 33, however, CF was significantly lower in the thermal stress group. The gill showed blood congestion in primary lamellae and shortened secondary lamellae in fish kept at 37–40 °C. No specific alterations were found in the cardiac tissue of fish kept in thermal stress conditions. Under thermal stress, 40% of fish survived until the end of the experiment. A preliminary conclusion drawn from this work is that A. dispar, which lives at normal temperatures, shows evidence of adaptability to elevated temperatures that could be a factor in the ability of killifish to adapt permanently over time to thermal environment.     

Influence of some environmental variables on the ascorbic acid status of mullet, Mugil cephalus L., tissues

The seasonal fluctuations in the ascorbic acid (AsA) and ascorbic acid 2-sulphate (AsA 2-sulphate) content of mullet, Mugil cephalus , tissues were examined. Ascorbic acid concentrations in brain, gill and hepatic tissues showed seasonal changes, but the pattern of AsA fluctuations in each tissue differed. The AsA content of mullet brains decreased during the summer, whereas hepatic AsA concentrations increased during this period and were maximal by the end of June. Hepatic AsA reserves declined after environmental water temperatures dropped below 18°C and reached a minimum (20 μg g⁻¹) by the end of January. Greatest fluctuations in AsA content occurred in gill tissues, which had a four-fold range of tissue concentrations. There were also seasonal changes in the AsA 2-sulphate content of brain and hepatic tissues. These differences among mullet tissues in the seasonal patterns of AsA content may be due to diverse effects of environmental variables on tissue AsA reserves. The ability of hepatic and renal tissues of mullet and several other teleost species to synthesize AsA was also investigated. L-gulonolactone oxidase activity was detected in all the species examined, but in all cases the biosynthetic capacity was less than a seventh that in goldfish, Carassius auratus , livers. Mullet appear to have only a limited capacity to synthesize AsA.     

Ethological and haematological response of catfish,Heteropneustes fossilis,exposed to exogenous urea

Exposure of the airbreathing catfish,Heteropneustes fossilis to a sublethal concentration (200 mg/l) of urea resulted in blanching of skin, increased mucus secretion, hyperexcitability and higher rate of surfacing and gill ventilation. These changes were more pronounced during the first eight days of exposure as compared to the later period. A progressive increase in haematocrit, erythrocyte count, percentage of immature erythrocytes, mean corpuscular volume and erythrocyte sedimentation rate was observed during the 20 day exposure period. White blood cell-thrombocyte count registered a sharp decline initially upto 6 days but steadily increased afterwards. Initial decrease in the total leucocyte count was found to be mainly due to the decline in the number of lymphocytes and thrombocytes while the percentage of granulocytes (neutrophils and eosinophils) and monocytes showed a reverse trend.     

Ameliorating effect of β-carotene on ethylmethane sulphonate-induced genotoxicity in the fish Oreochromis mossambicus

Genotoxic effects have been assessed in the fish Oreochromis mossambicus treated separately and conjointly with 0.2% ethylmethane sulphonate (EMS) and 0.05% β-carotene (BC) during five different time periods (6, 24, 48, 72 and 96 h) by analysis of endpoints such as chromosome aberrations, abnormal red blood cell nuclei, abnormal sperm morphology and protein contents (both qualitative and quantitative) of selected tissues, viz. muscle, heart, eye, brain, gill, liver, spleen and kidney. In addition, the relative efficacy of three doses of BC 0.02, 0.05 and 0.1%, in ameliorating genotoxic effects of 0.2% EMS was also tested after a treatment period of 48 h. EMS caused chromosomal aberrations, nuclear anomalies in red blood cells, abnormal sperm morphology and an apparent alteration of protein synthesis in various tissues. Some of these genotoxic effects of EMS appeared to be ameliorated by all three doses of BC, of which the 0.02% dose showed a marginally better efficacy.     

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.     

Adaptation to salinity by fish. Macromolecular changes in mitochondria and microsomes of the gill

Mitochondria isolated from the gill tissues of the fishSarothredon mossambica were analysed for their macromolecular content, following transfer from freshwater to media of higher salinity. The results suggest a breakdown of mitochondria during the initial phases of the stress and a regeneration during continued exposure. Also all the synthetic machineries, in general, seem to be triggered in gill tissue during continuous exposure to hyperosmotic media.     

Changes of gill and hemocyte-related bio-indicators during long term maintenance of the vent mussel Bathymodiolus azoricus held in aquaria at atmospheric pressure

The deep-sea hydrothermal vent mussel Bathymodiolus azoricus has been the subject of several studies aimed at understanding the physiological adaptations that vent animals have developed in order to cope with the particular physical and chemical conditions of hydrothermal environments. In spite of reports describing successful procedures to maintain vent mussels under laboratory conditions at atmospheric pressure, few studies have described the mussel's physiological state after a long period in aquaria. In the present study, we investigate changes in mucocytes and hemocytes in B. azoricus over the course of several months after deep-sea retrieval. The visualization of granules of mucopolysaccharide or glycoprotein was made possible through their inherent auto-fluorescent property and the Alcian blue-Periodic Acid Schiff staining method. The density and distribution of droplets of mucus-like granules was observed at the ventral end of lamellae during acclimatization period. The mucus-like granules were greatly reduced after 3 months and nearly absent after 6 months of aquarium conditions. Additionally, we examined the depletion of endosymbiont bacteria from gill tissues, which typically occurs within a few weeks in sea water under laboratory conditions. The physiological state of B. azoricus after 6 months of acclimatization was also examined by means of phagocytosis assays using hemocytes. Hemocytes from mussels held in aquaria up to 6 months were still capable of phagocytosis but to a lesser extent when compared to the number of ingested yeast particles per phagocytic hemocytes from freshly collected vent mussels. We suggest that the changes in gill mucopolysaccharides and hemocyte glycoproteins, the endosymbiont abundance in gill tissues and phagocytosis are useful health criteria to assess long term maintenance of B. azoricus in aquaria. Furthermore, the laboratory set up to which vent mussels were acclimatized is an applicable system to study physiological reactions such as hemocyte immunocompetence even in the absence of the high hydrostatic pressure found at deep-sea vent sites.     

Growth of the first antler in Iberian red deer (<Emphasis Type="Italic">Cervus elaphus hispanicus</Emphasis>)

In this study, we describe the process of pedicle and first antler growth in Iberian red deer (Cervus elaphus hispanicus) and document relationships among body development, maternal milk supply and composition, and maternal weight on the length of first antlers. Antler length of 53 males of Iberian red deer was measured every 2 weeks from birth to 20 months of age. Deer weight, age, and the date of occurrence of the major events during the antler growth cycle were also recorded. The first evidence of pedicle development occurred when the animals were 38.0 ± 0.6 weeks old and weighed 60.7 ± 0.9 kg. Antler cleaning took place at a mean age of 63.8 ± 0.7 weeks and a mean weight of 91.5 ± 1.8 kg. The antler growth period lasted 16.7 ± 0.4 weeks, and the cleaning period lasted 5.1 ± 0.4 weeks. First antler growth followed a sigmoid curve, reaching a final length of 38.3 ± 1.0 cm. Antler length was positively correlated with body weight during the antler growth cycle. Additionally, the final length of the first antler was related to total milk yield, date of antler growth initiation, body weight at 6 months of age, and the antler growth time interval.     

Multiple biomarker response in rainbow trout during exposure to hexavalent chromium

This research investigated hexavalent chromium toxicity in rainbow trout using a panel of biomarkers at different levels of biological organization. A time-course experiment in which rainbow trout were exposed in hard water (63.5 mg/L CaCO3) to a sublethal concentration of hexavalent chromium (10 mg/L) for a period of 28 days was conducted. The responses of multiple biomarkers were measured in gill and liver tissues at varying time points. Significant differences in metallothionein induction, superoxide dismutase activity, lipid peroxidation, cellular morphology, and growth were observed. Results indicated that gill tissues were more sensitive than hepatic tissues to chromium toxicity, yet hepatic tissues appeared to play a larger role in the organism's adaptive response to chromium compared to gill tissues. This study highlights the importance of using a set of integrated biomarkers to assess contaminant exposure and effects.     

Glucose-6-phosphate dehydrogenase activity in individual rat hepatocytes of different ploidy classes. I. Developments during postnatal growth

The glucose-6-phosphate dehydrogenase (G6PDH) activity of isolated male rat hepatocytes has been investigated in relationship to the ploidy classes of the cells during the first 20 weeks of postnatal growth. The G6PDH activity in the individual cells was measured with an improved quantitative cytochemical method. The data obtained showed that throughout the whole period of postnatal growth there existed a proportional relationship between the genome copies per cell and the amount of G6PDH activity per cell for binuclear diploid (BD), mononuclear tetraploid (MT) and binuclear tetraploid (BT) cells but not for mononuclear diploid (MD) cells. In the MD cells, which are the stem cells of the liver parenchyma, the activity measured was 1.5 times higher than expected. Furthermore, during postnatal growth, the G6PDH activity per hepatocyte was low at the age of 2 weeks, increased somewhat after weaning (5 weeks) and then more dramatically after 8 weeks to reach a maximum between 12 and 16 weeks. This development occurred in MT and BT cells at an earlier age than in MD and BD cells, in which the increase in enzyme activity followed some 3 weeks later. Castration of the rats before puberty did not influence the development of the amount of G6PDH activity per cell of any of the ploidy classes.     

Localization of the bioluminescence system in the pileus of Mycena chlorophos

Mycena chlorophos, which is primarily distributed in Southeast Asia, is a luminous fungus that emits a bright green light from its pileus for about 2 days at approximately 20°C and high relative humidity. The distribution of bioluminescent tissues in the whole pileus and its sections was heterogeneous. The light intensity in the cap and the upper region of the gill was greater than that in the lower region of the gill. At the microscopic level, the light was predominantly emitted from the membranes of hymenium and basidia cells on the gill. The emission was both cell and region specific. The luminescence system was localized in the cell membrane, and a part of the system was on the cell membrane surface. Copyright © 2015 John Wiley & Sons, Ltd.     

Anadromous trout threatened by whitefish gill‐net fisheries in the northern Baltic Sea

We estimated the effect of the gill‐net fisheries targeted at whitefish (Coregonus sp.) on anadromous sea trout, Salmo trutta, in the Gulf of Bothnia, Baltic Sea using separate data for fish species. The analysis of sea trout captures was based on tagging and recapture data collected in 1998–2011, while whitefish data were derived from individual samples of commercial fisheries from the same period. The mesh sizes used in gill‐net fishing and the seasonal and temporal distributions of recaptured sea trout and sampled whitefish were compared in the northern and southern Gulf of Bothnia. The trout had typically spent 1–2 years at sea, and they were mainly immature with a median body length of 40–43 cm at the time of recapture in gill nets. Despite the increase in the minimum permitted landing size from 40 to 50 cm in 2008, the median length of recaptured trout remained unchanged during the study period. Most (59%) of the gillnetted trout were caught in the southern Gulf of Bothnia in gill nets with mesh sizes of 40–45 mm, which were also used in the whitefish fishery (72%). In the northern Gulf of Bothnia, nets with a smaller mesh size of 25–39 mm took 83% of the whitefish catch and 39% from recaptured trout. In both areas, the overlap in mesh sizes used to gill‐net catch whitefish and sea trout increased during the study period. There were clear seasonal and areal differences in the relative probability of sea trout being captured in gill nets, suggesting that carefully tailored spatial and temporal restrictions on gill‐net fisheries could provide a tool to protect young sea trout without causing intolerable difficulties for the fisheries targeting other species.     

Variation in relative growth rate of 20 Aegilops species (Poaceae) in the field: The importance of net assimilation rate or specific leaf area depends on the time scale

Field experiments reporting the relative growth rate (RGR) patterns in plants are scarce. In this study, 22 herbaceous species (20 Aegilops species, Amblyopyrum muticum and Triticum aestivum) were grown under field conditions to assess their RGR, and to find out if the differences in RGR amongst species were explained by morphological or physiological traits. Plants were cultivated during two months, and five harvests (every 13–19 days) were carried out. Factors explaining between-species differences in RGR varied, depending on whether short (13–19 days) or longer periods (62 days) were considered. RGR for short periods (4 growth periods of 13–19 days each) showed a positive correlation with net assimilation rate (NAR), but there was no significant correlation with leaf area ratio (LAR) (with the exception of the first growth period). In contrast, when growth was investigated over two months, RGR was positively correlated with morphological traits (LAR, and specific leaf area, SLA), but not with physiological traits (NAR). A possible explanation for these contrasting results is that during short growth periods, NAR exhibited strong variations possibly caused by the variable field conditions, and, consequently NAR mainly determined RGR. In contrast, during a longer growth period (62 days) the importance of NAR was not apparent (there was no significant correlation between RGR and NAR), while allocation traits, such as LAR and SLA, became most relevant.     

Abscisic acid content at defined levels of bud dormancy and frost tolerance in two contrasting populations of Picea abies grown in a phytotron

Seedlings of a southern (Romanian) and a northern (Swedish) population of Picea abies were cultivated under continuous light and 20°C for 10 weeks. To arrest growth, induce terminal bud dormancy and promote frost tolerance the seedlings were then exposed to 16 h nights for 12 weeks, with gradually lower temperature during the last 6 weeks. Samples for estimating the abscisic acid content of the needles were taken just before the onset of the night treatment, at day 3 of the treatment, and then with one, and later 2 week, intervals. From the second week onwards (third week for frost tolerance) bud dormancy and frost tolerance were assessed at the same time as abscisic acid (ABA) determinations. Phosphate-buffered saline extracts were purified on mini-columns (in some cases immunoaffinity colums) and quantified by HPLC. The degree of dormancy was estimated by transferring the seedlings to growth conditions and determining the number of days until growth was resumed. The frost tolerance of the needles exposed to –10°C and –20°C was classified in 6 classes. The frost tolerance of the terminal buds was estimated as the number of seedlings that showed some growth after 6 weeks in growth conditions. The night treatment rapidly induced terminal bud dormancy in both populations, but the release of dormancy occurred earlier in the northern population. The needles and the terminal buds became highly frost tolerant more rapidly in the northern than in the southern population and before the temperature decrease. The degree of dormancy began to decline before full frost tolerance was obtained in the southern population and this decline continued in both populations, while frost tolerance remained at a high level. The southern population showed a transient peak in ABA content at day 3. Although the ABA content of the northern population was lower than in the southern before the 16-h night treatment, it increased in the northern population during the treatment period, in particular after the temperature decrease.     

Elevated spring temperature stimulates growth, but not smolt development, in anadromous Arctic charr

Parr–smolt transformation and growth were studied in captive offspring of anadromous Arctic charr (Salvelinus alpinus) from the Hals watercourse in northern Norway (70°N), held either at a natural temperature (< 1 °C until May) or at a temperature elevated to 6 °C in late March. In mid-May, 5 weeks after the increase in photoperiod from 8:16 h light:dark to continuous light, gill Na⁺, K⁺-ATPase activity started to increase in both temperature groups, concurrent with the final development of full seawater tolerance. Temperature had no effect on the development of gill Na⁺, K⁺-ATPase activity, or on hypoosmoregulatory ability. The fish in both treatments resumed growth in mid-May, but from then on growth was faster in the elevated than in the ambient temperature group. In the former group, fish mass doubled in 6 weeks (from 65 to 137 g), and growth ceased at the time when the fish were about to complete their parr–smolt transformation. These findings show that an early vernal temperature increase advances the seasonal growth cycle, but not the parr–smolt transformation, in anadromous Arctic charr.     

Sublethal effects of an organophosphorus insecticide (RPR-II) on biochemical parameters of tilapia, Oreochromis mossambicus

The effect of exposure to sublethal concentrations (0.017 mg L(-1), 1/10 of LC50) of the novel organophosphate (OP) insecticide, 2-butenoic acid-3-(diethoxyphosphinothioyl) methyl ester (RPR-II) on biochemical parameters in Oreochromis mossambicus was studied during exposure for 3, 7, 15, 30 and its recovery response after seven days. Acetylcholinesterase (AChE) activity of brain, gill and muscle was inhibited by 67%, 77% and 73% respectively on day-30. The plasma and kidney alanine aminotransferase (AlaAT), and aspartate aminotransferase (AspAT) activity increased, while decreases were observed in gill and liver. Increases in acid phosphatase (AcP), and alkaline phosphatase (AP) activities were observed in plasma, gill, and kidney, and reductions of 20% and 61% in liver AcP and AP, respectively. Depletion of glycogen was observed in all tissues, an indication of typical stress related response of the fish with pesticide. Lactate dehydrogenase (LDH) activity decreased in liver and muscle, indicating tissue damage but a significant increase in LDH activity in gill and brain was observed. Depletion of glutathione (GSH) was observed in all tissues, thereby enhancing lipid peroxidation resulting in cell damage. The induction in hepatic glutathione-S-transferase (GST) levels indicates protection against the toxicity of xenobiotic-induced lipid peroxidation. There was a significant recovery in the above biochemical parameters, in all tissues of fish after a recovery period of seven days. These results revealed that the OP insecticide RPR-II is highly toxic and affects the intermediary metabolism of O. mossambicus.