Effect of cadmium on ciliary and ATPase activity in the gills of freshwater mussel Anodonta cygnea

1. Cadmium (≤ 50 μM) decreases the heat resistance (39°C) of the activity of frontal cilia in the Anodonta cygnea gills incubated in dechlorinated tap water, while in the presence of added 2 mM Ca²⁺ the minimal acting concentration of cadmium rises up to 100 μM.2. The inhibitory effect of Cd²⁺ (1.5 mM) on the ATPase activity measured in the gill microsomal fraction is temperature dependent and increases as follows: ouabain insensitive Na²⁺- or K⁺-ATPase (no inhibition), Ca²⁺-ATPase (50% inhibition), Mg²⁺-ATPase (100% inhibition).3. Cadmium itself (≤ 50 μM) added to microsomal suspension stimulates the H⁺-sensitive ATP hydrolysis resembling on its pH-dependence the Mg²⁺- but not Ca²⁺-ATPase activity.4. Cd²⁺ can mimic the effect of Mg²⁺ as a cofactor required for activation of the ouabain-insensitive Na⁺- or K⁺-ATPase. Monovalent cations fail to activate the ATPase when Mg²⁺ is substituted by Ca²⁺.5. One of the mechanisms underlying the toxicity of Cd²⁺ to Anodonta gills could be based upon an interaction of Cd²⁺ with Mg²⁺-ATPase followed by suppression of the ciliary activity.     

Effect of sub-lethal concentrations of aluminium on the filtration activity of the freshwater mussel Anodonta cygnea L. at neutral pH

Significant amounts of aluminium (Al) are commonly present in rivers and lakes, largely in particulate form in neutral waters. Freshwater bivalves, as filter feeders are therefore exposed to both particulate and dissolved metal and are potentially vulnerable to Al. The effect of Al on filtering behaviour of the freshwater mussel Anodonta cygnea L. was investigated during short (1 hour) and long-term (15 days) exposure to environmentally relevant concentrations (250 and 500 microg l(-1)) at neutral pH. Water flow through the outflow siphon was monitored as an indicator of pumping capacity. Short-term (1 hour) exposure to 500 microg l(-1) added Al produced an irreversible decrease in the duration of filtering periods, presumably as an avoidance response to the toxicant. One-hour exposure 250 microg l(-1) Al had no detectable effect. When mussels were exposed to 250 or 500 microg l(-1) added Al for 15 days, siphon activity measured in days 11-15 of exposure was inhibited by 50% and 65%, respectively, compared to pre-exposure levels. Recovery occurred following transfer of mussels to uncontaminated water. Interaction between Al and freshwater bivalves at neutral pH may affect both the performance of the mussels and the chemical speciation of the metal in the natural environment.     

Mineral composition variation in the shells of freshwater mussel Anodonta woodiana at different growth stages

The levels of mineral element Na, Mg, K, Ca, Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, Mo, Al, As, Ag, Cd, and Tl were quantified in the whole shells of the freshwater bivalve Anodonta woodiana at three different growth stages (i.e. J1 juveniles of 1 month old, J2 juveniles of 3.5 months old, and adults of 36 months old). The concentrations of Na and Al were different between different growth stages (p < 0.05). The highest Na concentrations (2715 ± 86 μg/g dry weight) were found in J2 juveniles. The highest Al concentrations (303.9 ± 5.95 μg/g dry weight) were found in J1 juveniles. Manganese concentrations (517.0 ± 47.98 μg/g dry weight) were significantly higher in J2 juveniles than in J1 juveniles (432.3 ± 9.87 μg/g dry weight) (p < 0.05). Copper concentrations (27.32 ± 0.15 μg/g dry weight) were significantly higher in J1 juveniles than in J2 juveniles (26.21 ± 0.86 μg/g dry weight) and adults (24.74 ± 1.43 μg/g dry weight) (p < 0.05). Burdens of Na, Ca, Mn, Fe, Co, Cu, Mo, Ag, and Tl were positively correlated with the shell length (p < 0.05). These findings can possibly contribute to an understanding of elemental requirements for shell growth and, hence, facilitate improvement of survival and growth rates during artificial mussel culture.     

Growth and age structure of the swan mussel Anodonta cygnea (L.) at different depths in lake Mattsee (Salzburg, Austria)

Unionid clams were collected at 1–2 m, 3–4 m and 6–7 m depth in lake Mattsee, a moderately mesotrophic lake, to investigate the effect of depth on clam growth and age structure. No significant differences in age structure of Anodonta cygnea were found (p=0.65). Three and ten years old clams were present at all depths, but in different percentages. Whereas at 1–2 m 13.3% of the collected clams were <4 years old, this percentage was 4.4% at 6–7 m and 7.1% at 3–4 m. A greater percentage (6.7%) of older mussels (9, 10 years) were collected at 6–7 m than at 1–2 m (2.2%). Growth declined with depth. Total length at a given age of clams at 1–2 m and 3–4 m did not differ (p=0.54), whereas differences were significant between clams at 1–2 m and 6–7 m (p<0.05) as well as between 3–4 m and 6–7 m (p<0.05). The Growth constant k was highest at 1–2 m depth.     

Reproduction and larval developmental stages of the freshwater mussel Limnoperna fortunei at three sites in a temperate reservoir

Longitudinal changes in larval densities and developmental stage compositions of the freshwater mussel Limnoperna fortunei were investigated to understand the patterns of reproduction and development at three sites in Lake Ohshio, a small reservoir in the temperate region of central Japan. Onset and termination of L. fortunei larval presence were coincident at the three sites in the reservoir. This may have been caused by environmental homogenization owing to artificial aeration for water quality improvement of the reservoir. The observations in this study support temperature as the trigger for onset of reproduction as suggested by previous studies. Occurrence of high turbidity that reduced food availability for mussels was probably the trigger for termination of reproduction in Lake Ohshio. During the period when L. fortunei larvae were present, the D-form larva of the earliest developmental stage was always observed and at much higher density than later developmental stages. The results indicate that spawning of L. fortunei continued in the reservoir throughout the period that larvae were observed and that predation and other factors reduced the proportion of later developmental stage larvae.     

The neuropeptide TRH has a minor effect on the enzymatic activity of acetylcholinesterase in vitro

The neuropeptide thyrotropin-releasing hormone (TRH) elicits a variety of physiological effects of which some are due to cholinergic mechanisms. TRH modulates in vivo the effects of compounds affecting acetylcholinesterase (AChE). In the present study the in vitro effects of TRH on the activity of AChE were explored. TRH has no effect at physiologically relevant concentrations. At unphysiologically high concentrations (>5 mM) a slight inhibition was found. This was noticed also when the enzyme was exposed to the amide-free tripeptide analog p-Glu-His-Pro. We conclude that any cholinergic effect of TRH observed in vivo is unlikely to be due to a direct interaction of the peptide with AChE.     

Measuring the impact of sewage effluent on the macroinvertebrate community of an upland stream: The effect of different levels of taxonomic resolution and quantification

Abstract Benthic macroinvertebrates were sampled from four sites on upland streams in the Wentworth Falls area of the Blue Mountains, NSW. One site received effluent from a sewage treatment plant and the others were reference sites. Five replicate collections were taken from each site on four occasions at intervals of 3 months. Macroinvertebrate community data were analysed using univariate (ANOVA) and multivariate (NMDS) techniques and comparisons were made between analyses at different levels of taxonomic aggregation and using different methods of data transformation. Similar patterns were observed at both species and family levels, and even the order level showed a clear community response to effluent input. Binary (presence/ absence) data provided similar results to quantitative data for the species and family levels. However, when binary data were used at the order level, the distinctions between the reference sites became blurred. We discuss the implications of these findings for environmental monitoring.     

Fine-structural localization of proenkephalin mRNAs in the hypothalamic magnocellular dorsal nucleus of the guinea pig: a comparison of radioisotopic and enzymatic in situ hybridization methods at the light- and electron-microscopic levels

With the aim of localizing proenkephalin mRNAs in neurons of the hypothalamic magnocellular dorsal nucleus of the guinea pig, we compared the in situ hybridization signals obtained on Vibratome sections with a method employing either a biotinylated or a digoxigenin-labeled oligonucleotide detected by means of the alkaline phosphatase reaction. Since the hybridization approach using the biotinylated probe was more sensitive than the digoxigenin method, the ultrastructural localization of hybrids in neurons of the magnocellular dorsal nucleus was studied by the use of the former procedure, and was further compared with results of in situ hybridization using a ³⁵S-labeled probe. Biotin was detected via an amplified avidin-biotin-peroxidase complex. Radioactive hybrids were localized over extended cytoplasmic compartments rich in rough endopoasmic reticulum and also in nuclear indentations. The method based on biotinylated probe proved to be sensitive and provided high-resolution labeling in well-preserved specimens. Proenkephalin mRNAs were clearly localized within circumscribed cytoplasmic compartments. The immunoprecipitates were mainly observed within the rough endoplasmic reticulum, especially at the periphery of the cell. The reticulum was dominated by elongated parallel cisternae. The labeling also appeared in a paranuclear position, mainly in nuclear indentations. The labeling was found on the outer surface of the endoplasmic lamellae. The remainder of the reticulum was unlabeled. Neuronal processes were free of labeling.     

Effects of municipal effluents on serotonin and dopamine levels in the freshwater mussel Elliptio complanata

Sex differentiation and gametogenesis represent critical steps in the reproductive process and are subject to hormonal control by serotonin, dopamine and steroids such as estradiol-17beta and testosterone. The purpose of this study sought to examine the endocrine-disrupting activity that a primary-treated municipal effluent might have on the metabolism of biogenic amine levels. First, serotonin receptors transfected in Chinese hamster ovary (CHO) cells were used to screen for the presence of serotonin receptor agonist or antagonist. Second, one group of Elliptio complanata mussels were exposed to single compounds likely to be found in municipal wastewaters and another group was exposed in situ to the municipal effluent plume for 90 days in experimental cages. Results showed that solid phase C-8 extracts of surface water downstream a municipal effluent could activate the transport of serotonin by receptors at a distance of at least 5 km from its outfall thereby indicating the presence of serotonin mimics in the effluent dispersion plume. Levels of serotonin and monoamine oxidase (MAO) activity in nerve ganglia of mussels exposed for 90 days to the municipal effluent were, respectively, reduced and increased at a distance 10-km downstream. Injections of estradiol-17beta and nonylphenol in mussels decreased the levels of serotonin and dopamine, but increased MAO activity in the gonad and nerve ganglia. Exposure to estrogenic chemicals present in municipal effluents may therefore alter the normal metabolism of serotonin and dopamine, both of which are involved in sexual differentiation in bivalves and fish. Chemicals acting through E2 receptor-mediated pathways and serotonin receptors are likely to cause the observed effects.     

Fluoride exposure and factors affecting dental caries in preschool children living in two areas with different natural levels of fluorides

BackgroundSuccessful oral health interventions must be based on the specific needs of the population that they serve. Evaluation of habits related to dental caries development and estimation of fluoride exposure in a target group of young patients helps to plan effective and safe caries prevention strategies.ObjectivesThe study aimed to evaluate factors affecting dental caries experience and sources of fluoride exposure in preschool children living in two areas: with optimal and low natural content of fluoride in drinking water.Materials and methodsThe study included a group of 73 children of both sexes aged 4–7 years attending two kindergartens in Środa Wielkopolska and Turek (Wielkopolska Voivodeship, Poland), where the content of fluoride in drinking water according to data obtained in the sanitary station ranged from 0.68 to 0.74 mg/L (optimal concentration of fluoride) and from 0.19 to 0.30 mg/L (low concentration of fluoride), respectively. Parents of patients completed a survey about diet, hygiene, and dental care, taking into account the child's fluoride exposure. The calibrated dentist assessed the oral health condition using a mirror, a CPI probe, and a headlamp. Oral hygiene was recorded using the Silness and Löe plaque index, caries experience by calculating the numbers of decayed, missing, and filled primary, and permanent teeth (dmf and DMF, respectively) while caries frequency by calculating the percentage of children with caries experience above 0. In order to assess the fluoride concentrations in urine and drinking water, parents were asked to provide a urine sample collected on fasting and a tap water sample. Fluoride concentrations were assessed using a 09–37 (MARAT) fluoride ion-selective electrode and a RAE 111 silver-chloride reference electrode. Statistical analysis was conducted using the data analysis software system Statistica (version 12, StatSoft, Inc. 2014), assuming a statistical significance level p < 0.05.ResultsNo statistically significant differences were found between caries indices of the examined children in each kindergarten (p > 0.05). Urinary fluoride levels were higher in children who tended to swallow toothpaste or used fluoride rinses and positively correlated with fluoride concentrations in the drinking water. Dental caries experience in the examined children depended on the effectiveness and frequency of oral hygiene procedures and dietary habits.ConclusionsThe strategy aimed at improving the oral health of the examined group of children should include accomplishing oral hygiene, promoting a non-cariogenic diet, and, finally, controlling fluoride exposure from at-home fluoride products. Caries prevention program ought to be adjusted to individual characteristics of each child, taking into consideration oral hygiene practices, dietary habits and total fluoride intake.     

Effect of propranolol and cold exposure on the activities of antioxidant enzymes in the brain of rats adapted to different ambient temperatures

The exposure to cold (6 h; 6°C) induced a significant decrease in both hypothalamic and brain stem CuZn-superoxide dismutase as well as an increase in Mn-superoxide dismutase and catalase activities in Wistar male rats, acclimated to 6±1°C as compared to those acclimated to 22±2°C. If the rats were administered with propranolol (15 mg/kg), which is a β-adrenoceptor blocker, there were no significant differences in the enzyme activities in any of the brain regions of the two groups studied. It was concluded that acute exposure to cold induces changes in the hypothalamic and brain stem antioxidant enzyme activities dependent on the previous acclimation to different ambient temperatures and propranolol administration.     

不同高程短尖苔草对水位变化的生长及繁殖响应

在淡水湿地生态系统中,水位通常是制约植被生长和繁殖动态的关键因素,进而对物种组成、群落演替和植被分布格局产生决定性影响。无性繁殖是洞庭湖湿地克隆植物适应环境胁迫的重要策略之一,以洞庭湖湿地典型克隆植物-短尖苔草(Carex brevicuspis C.B.Clarke)为对象,研究了不同分布高程(23.7 m和25.8 m)的植物对水位变化(0 cm,-15 cm,-30 cm)的生长和繁殖特征响应。结果表明:水位变化对不同分布高程分布短尖苔草的生长和克隆繁殖特征均产生显著影响(P0.05)。对高程区的短尖苔草而言,克隆繁殖特征如分株数、分株总生物量、芽数和芽生物量随水位的降低而增加,而对生长特征(株高及总生物量)无显著影响(P0.05),表明适当干旱有利于高程区苔草的克隆繁殖。对于低程区分布短尖苔草而言,水位变化对其生长特征有显著影响(P0.05),如株高和总生物量随着水位的降低而增加;分株数和总芽生物量等克隆繁殖特征则随水位的降低而减少,而水位对低程区短尖苔草的分株总生物量和总芽数影响不显著(P0.05)。因此,短尖苔草的克隆繁殖特征除受到水位的影响外,还受其分布高程的影响。可见,同一种短尖苔草因长期适应于不同生境而对相同的环境胁迫表现出了不同的生长繁殖策略,     

The role of biotic and abiotic factors in evolution of ant dispersal in the milkwort family (polygalaceae)

A phylogenetic approach was taken to investigate the evolutionary history of seed appendages in the plant family Polygalaceae (Fabales) and determine which factors might be associated with evolution of elaiosomes through comparisons to abiotic (climate) and biotic (ant species number and abundance) timelines. Molecular datasets from three plastid regions representing 160 species were used to reconstruct a phylogenetic tree of the order Fabales, focusing on Polygalaceae. Bayesian dating methods were used to estimate the age of the appearance of ant-dispersed elaiosomes in Polygalaceae, shown by likelihood optimizations to have a single origin in the family. Topology-based tests indicated a diversification rate shift associated with appearance of caruncular elaiosomes. We show that evolution of the caruncular elaiosome type currently associated with ant dispersal occurred 54.0-50.5 million year ago. This is long after an estimated increase in ant lineages in the Late Cretaceous based on molecular studies, but broadly concomitant with increasing global temperatures culminating in the Late Paleocene-Early Eocene thermal maxima. These results suggest that although most major ant clades were present when elaiosomes appeared, the environmental significance of elaiosomes may have been an important factor in success of elaiosome-bearing lineages. Ecological abundance of ants is perhaps more important than lineage numbers in determining significance of ant dispersal. Thus, our observation that elaiosomes predate increased ecological abundance of ants inferred from amber deposits could be indicative of an initial abiotic environmental function.     

The effect of feedback regulation and in situ product removal on the conversion of sugar to cycloheximide by Streptomyces griseus

An addition of cycloheximide to cycloheximide-producing Streptomyces griseus cultures resulted in reductions in the production rate and in the conversion of sugar into cycloheximide. In situ cycloheximide adsorption was observed to enhance: total cycloheximide titers; productivities; and the conversion of sugar to cycloheximide. During the secondary metabolite-producing phase, sugar consumption was observed to be linearly dependent on cycloheximide productivity. From this analysis a true product yield and maintenance coefficient were estimated to be 0.08 g cycloheximide/g glucose and 0.028 g glucose/g cell-h, respectively. The sixfold difference between this true product yield and a theoretical value obtained from knowledge of the biosynthetic pathway is discussed. Since the maintenance sugar requirement for cycloheximide production is large, stimulation of biosynthesis through in situ adsorption significantly increases the overall efficiency of sugar conversion to this secondary metabolite.     

Effects of different abiotic and biotic factors on spatial primary seed dispersal in the semachorous species Scrophularia canina

Seed dispersal is one of the most important steps in the plant life cycle. However, there is, generally, a lack of fieldworks focused on wind dispersal and especially on semachorous dispersal (seeds spread when the fruits are shaken by wind and other vectors, such as animals), including boleochorous dispersal. Therefore, we aimed to determine how different types of wind and animals affected seed dispersal under natural conditions in the widespread species Scrophularia canina. We evaluated the effects of wind gusts (simulating them using a leaf blower) and wild animals (using differently sized dogs) on seed dispersal in a population located in south-western Europe. We found that S. canina is a semachorous species, and its spatial seed dispersal was affected by wind gust speed and direction, plant structure and vector type. The results also revealed the presence of xerochasy, individual anisotropy with strong winds, and primary short-distance dispersal associated with successional processes independent of the vector. Additionally, there was a masking effect of plant structure on the seed shadow outline. It is essential to conduct fieldworks to reveal what actually happens in nature, taking into account the characteristics determining seed dispersal. In addition, in these works it is important to find out what factors affect seed distributions of anemochorous and semachorous species.     

Application of a direct toxicity assessment approach to assess the hazard of potential pesticide exposure at selected sites on the Crocodile and Magalies rivers,South Africa

The potentially hazardous effects of agricultural pesticide usage in the Crocodile (west) Marico catchment were evaluated using the Danio rerio and Daphnia pulex lethality, Selenastrum capricornutum growth inhibition and the Ames mutagenicity plate incorporation assays. Hazard assessment categories are proposed to standardise the output of the different toxicity assessments. Sites were categorised according to the toxicity hazard indicated and were related to the gradients of agricultural land-use intensity at each site. Intensive agricultural sites showed the highest effects to all tested biota. Receiving water at urban sites associated with increased nutrients and lowest pesticide usage showed few adverse effects, while the relatively unimpacted site indicated no hazard to any organism, and only a slight stimulation to algal growth. Weighted hazard scores indicated that the unimpacted sites were least hazardous, falling within a B category, the urban sites were moderately hazardous (C category), and the agricultural sites (D category) had the highest potential impacts on aquatic organisms. This study demonstrated the usefulness of using the hazard assessment approach and the role it could play in assessing site-specific potential toxicity hazards of river water impacted by agrochemicals. It can be used together with other assessment methods, such as biological indices, in a tiered approach.     

Effects of parasitism and pesticide exposure on characteristics and functions of hemocyte populations in the freshwater snail Lymnaea palustris (Gastropoda, Pulmonata)

Morphological characteristics and functions of hemocytes were used to compare the immunological effects of biological and chemical stress in the freshwater snailLymnaea palustris. Animals were either infected by a trematode parasite (Metaleptocephalus sp.), or exposed to environmental contaminants, namely atrazine and hexachlorobenzene (HCB). Three populations of circulating hemocytes, morphologically and cytochemically distinct (round cells, hyalinocytes, granulocytes), were identified in both control and parasitized or pesticide-exposed snails. After 6 h of exposure, HCB and atrazine resulted in 8-fold increases in the mean total number of hemocytes, whereas only a 2.2-fold increase was observed 6 h after cercaria emission in parasitized snails. The impact of HCB was limited to the first 24 h of exposure, whereas long-lasting effects of atrazine were observed. Hyalinocytes and, to a lesser extent, round cells contributed most to the increases in hemocyte density in pesticide-exposed snails. Parasitism and atrazine treatment resulted in significant increases of lectin-stained hemocytes, whereas exposure to HCB did not affect the percentages of stained and unstained cells. Hemocyte phagocytic activity increased in HCB-exposed snails but with no concomitant change of the oxidative burst. Opposite results were obtained in atrazine-treated snail hemocytes, with unchanged phagocytosis and decreased phorbol 12-myristate 13-acetate-stimulated production of reactive oxygen intermediates. No increase in phagocytosis, or in the production of reactive oxygen intermediates, was observed in hemocytes from parasitized snails. Infection with the immunologically compatible trematode parasiteMetaleptocephalus sp. and exposure to atrazine generated similar reactions from circulating hemocytes, whereas a different response pattern was observed in HCB-exposed snails. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effects of flow and stream characteristics on the variation in freshwater mussel growth in a Southeast US river basin

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