Activities of glutamate dehydrogenase and aspartate and alanine aminotransferases in freshwater snails Helisoma duryi and Lymnaea natalensis exposed to copper

In this paper we investigate the potential of glutamate dehydrogenase (GDH) and aspartate and alanine aminotransferases (AST and ALT) as biomarkers of water pollution due to copper in the freshwater snails Helisoma duryi and Lymnaea natalensis. Snails were dosed with copper(II) ion concentrations of 0.01, 0.1 and 1 mg kg^-1 breeding water for a period of 96 h, after which those surviving were shelled. The copper content in the breeding water, in whole snail tissue and in the snail shells was determined at the end of the period of exposure. For enzyme determinations, whole snail tissue was first homogenized and fractionated by centrifugation at 500 g to remove the nuclei. The resulting supernatant was then centrifuged at 10 000 g to give a pellet fraction representing the mitochondrial fraction and a supernatant representing the cytosolic fraction. Copper was very toxic to both snail species at concentrations above 0.2 mg l^-1, with only 3% of the Helisoma and 12% of the Lymnaea surviving at concentrations of approximately 1 mg l^-1. The copper content in the shells and tissues of snails rose with increasing copper concentration in the breeding water, and was 2.1- to 4.9-fold in snails exposed to copper ion at a dose of 1 mg kg^-1 water compared with undosed snails. Similarly, the activities of GDH and AST rose by up to 4.7-fold in the homogenate and the mitochondrial and cytosolic fractions with increasing concentrations of copper. These activities, however, fell at copper concentrations of approximately 1 mg l^-1, which coincided with massive death of snails. Mitochondrial ALT disappeared at copper ion concentrations of approximately 0.2 mg l^-1 for Lymnaea and 1 mg l^-1 for Helisoma, possibly indicating mitochondrial degeneration. These results show that GDH, AST and ALT have the potential to be biomarkers of suplethal copper pollution in these two snail species, since their activities were significantly altered by low copper concentrations.     

提高实验猴繁殖率的综合措施

实验猴繁殖率的高低直接影响实验猴养殖企业的经济效益。本文借鉴本中心多年实验猴繁育经验,总结了一套较为完备的提高实验猴繁殖率的措施,内容涵盖实验猴繁殖特性、种公猴选种、繁殖猴分群及梯度饲养、孕猴用药、母猴产后护理、仔猴补饲及断奶、繁殖猴饲料搭配等各方面。这些措施应用于实践后,实验猴繁殖率可保持在较高水平并逐年提高。我们认为这些措施可在其它实验猴养殖企业推广应用。     

The influence of dietary copper on reproduction, growth and the cardiovascular system in Swiss-Webster female mice

One hundred forty pubertal Swiss-Webster female mice (Mus musculus) were assigned randomly to a purified diet modified to contain 1, 2, 3, 4, 5 and 6 ppm copper as copper carbonate (Diets 1 thru 6, respectively). Diet 7 was a commercial rodent laboratory diet containing 11 ppm copper. Following 60 days of prefeeding, the breeding regimen was initiated by introducing a proven breeder male into each cage of 10 females in the afternoon and removing them the next morning. The breeding regimen was conducted for 28 days and females were either sacrificed at 24 or 96 hours following detection of a vaginal plug. Body weight, hematocrit, heart weight, hemoglobin concentration, ova recovery rate and fertilization rate were recorded for each female. All morphologically normal embryos collected 24 hours after detection of the vaginal plug were cultured and development in vitro evaluated. Body weight, hemoglobin concentration and hematocrit were lower (P less than .05), and heart weight and percent body weight occupied by the heart (%BWOH) were higher (P less than .05) in mice fed Diet 1. Heart weight and %BWOH were not different among mice fed Diets 2 thru 6 (P greater than .05). Ova recovery rate and fertilization rate were significantly reduced in mice maintained on Diet 1. The incidence of in vitro blastocyst formation was lower in embryos collected from females fed Diets 1 and 3 (P less than .05). Blastocyst hatching in vitro was not observed in embryos from females maintained on Diet 1, and was greater in embryos from females fed Diets 4, 5, and 7 (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

鱼体(去鳃)和鱼鳃对不同形态铜的积累特征

在实验室条件下研究实验鱼Ｐａｒａｃｈｅｉｒｏｄｏｎ对人工河水中不同形态的积累特征,对比了鱼体（去鳃）和鱼鳃对铜吸收量的差异,并探讨了鱼对铜的吸收机理。研究结果表明,实验鱼鳃部和体内铜积累量均随水相游离铜浓度增高,暴露时间增长而增加,但鳃部积累浓度较鱼体其余部分高一个数量级,其从水相富集铜的速率显著高于鱼体。     

铜污染对昆虫生长发育及繁殖影响的研究进展

铜(Cu)是生物体生长发育所必需的微量营养元素,参与或构成许多含铜酶及含铜生物活性蛋白质,但是过度摄入Cu会对生物体产生危害。Cu中毒由许多因素而定,如物种、遗传、年龄和食物等。在生态系统中昆虫种类多,数量大,分布广,往往会取食、转移和代谢环境中的Cu,从而对自身生长发育及繁殖造成影响。通常表现为发育历期延长,体重下降,体长变短,化蛹率、羽化率、产卵率、孵化率、存活率降低,种群数量减少等。本文以近几年Cu2+胁迫对昆虫生长发育及繁殖影响的研究进行综述。     

Breeding and growth of the inbred strain of seizure-prone Mongolian gerbils (Meriones unguiculatus)

Recently a Mongolian gerbil has become a useful laboratory animal, while little of the basic data are known about the breeding and growth of this laboratory rodent. In our animal center, an inbred strain of the seizure prone gerbil has been produced and kept these ten years. The data on the breeding and growth of the animal were obtained and described as following in the present paper. The mean life span was 26 +/- 14 month old for males and 27 +/- 13 month old for females. The mean body weight of the Mongolian gerbils at 16 weeks of age was 72 +/- 5.6 g for males and 62 +/- 7.3 g for females. The mean litter size at birth was 4.8 +/- 1.7 head. (range; 1-9) More newborn pups were born in the warm seasons (March-October) than the cold seasons (November-February). The mean rate of weaning was 80.2%. The man litter size was relatively constant in each generation.     

Effects of dairy wastewater on mosquitoes in southern California.

The effect of dairy pond water quality on mosquito biology was studied under laboratory conditions during 2004. Due to high turbidity and larval toxicity of raw water samples from dairy ponds with no mosquito breeding, water samples from both mosquito breeding and non-breeding ponds were filtered and diluted 5X with distilled water prior to being tested for sublethal effects on mosquito development. The diluted pond waters showed no significant deleterious effects on the overall development of Culex quiquefasciatus Say at 23.9 degrees and 26.6 degrees C. However, the 3rd and 4th instar larvae were significantly shorter and lighter in weight when reared in diluted water samples from mosquito non-breeding ponds. Similar effects were found on the developmental stages of Cx. tarsalis Coquillette. Data on water quality parameters of filtered pond waters showed higher levels of hexavalent chromium (4X), nitrites (10X), nitrates (2X), sulfates (5X), and salinity (2X) in water samples from non-breeding ponds. Based on these data, the presence of high levels of hexavalent chromium and sulfate appear to be responsible for the absence of mosquito breeding in some dairy ponds.     

Influence of breeding site availability on the oviposition behaviour of Aedes aegypti

Despite the importance of the mosquito Aedes aegypti in the transmission of arboviruses, such as yellow fever, Chikungunya fever and dengue fever, some aspects of their behaviour remain unknown. In the present study, the oviposition behaviour of Ae. aegypti females that were exposed to different densities of breeding sites (2, 4, 8 and 16) was evaluated in laboratory and semi-field conditions. The number of breeding sites that were used was proportional to the number available, but tended towards stabilisation. Females used four-six breeding sites on average, with a maximum of 11. A high percentage of eggs was observed in the water, along with the presence of a breeding site termed “favourite”, which received at least 40% of the eggs. The results are discussed in ecological, evolutionary and epidemiological approaches.     

Body condition dynamics and the cost‐of‐delay hypothesis in a temperate‐breeding duck

Pre‐breeding body condition is an important determinant of reproductive success in birds, largely through its influence on timing of breeding. Declines in clutch size and recruitment probability within breeding seasons indicate a tradeoff may exist between the number of young (clutch size) and quality of young (recruitment probability). We explored local drivers of pre‐breeding body condition and tested predictions of the cost‐of‐delay hypothesis in female lesser scaup Aythya affinis. Yearling females arrived on the study site in lower body condition than older females, but both age classes had similar rates of body condition gain on the breeding grounds prior to nesting. Rates of body condition gain were positively influenced by water temperature, a proxy for wetland phenology. The effect of water level was asymptotic and interacted with water temperature, with greater rates of gain in body condition occurring in years with low water levels. Our results supported the predicted response of clutch size to the rate of pre‐breeding body condition gain. After accounting for lay date, clutch size was positively related to the rate of body condition gain (= 0.08 ± 0.039). We did not find support for a predicted interaction between rate of body condition gain and intra‐seasonal decline in clutch size (= 0.01 ± 0.01). Our results indicate that local conditions during pre‐breeding influence body condition dynamics in female lesser scaup, which subsequently affects clutch size.     

Nesting area fidelity and survival of female Common Goldeneyes Bucephala clangula: are they density‐dependent?

The breeding histories of 218 female Common Goldeneyes Bucephala clangula were recorded between 1971 and 2000 in a study area in Schleswig-Holstein, northern Germany. Females were first recorded breeding at a median age of 2 years usually in their area of hatching (philopatry). One hundred and two of 140 females (73%) re-nested only in one of the 13 nesting areas (clusters of nestboxes) for all their known life of up to 13 breeding seasons. The remaining 38 individuals moved between different nesting areas at least once between breeding attempts. The two oldest females were still breeding at a minimum age of 15 years (i.e. 13 years between first and last recorded breeding attempt). Temporal variations in annual survival rates of adult females could best be explained by a model with annual survival rate varying independently and randomly about a mean of 0.830 (se = ±0.023) with an estimated sd of ±0.092 (95% CI = 0.064, 0.138). No trend in the annual survival rate was detected over the study period of 30 years, although the presence of a moderate trend could not be ruled out. The absence of any discernible trend in survival at a time when the population size increased substantially indicates little, if any, density-dependence in survival of female Goldeneyes during this study.     

Genetic Adaptation to Captivity and Inbreeding Depression in Small Laboratory Populations of Drosophila Melanogaster

The rate of adaptation to a competitive laboratory environment and the associated inbreeding depression in measures of reproductive fitness have been observed in populations of Drosophila melanogaster with mean effective breeding size of the order of 50 individuals. Two large wild-derived populations and a long-established laboratory cage population were used as base stocks, from which subpopulations were extracted and slowly inbred under crowded conditions over a period of 210 generations. Comparisons have been made of the competitive ability and reproductive fitness of these subpopulations, the panmictic populations produced from them by hybridization and random mating and the wild- or cage-base populations from which they were derived. After an average of ~180 generations in the laboratory, the wild-derived panmictic populations exceeded the resampled natural populations by 75% in fitness under competitive conditions. The cage-derived panmictic population, after a total of 17 years in the laboratory, showed a 90% superiority in competitive ability over the corresponding wild population. In the inbred lines derived from the wild-base stocks, the average rate of adaptation was estimated to be 0.33 +/- 0.06% per generation. However, the gain in competitive ability was more than offset by inbreeding depression at an initial rate of ~2% per generation. The effects of both adaptation and inbreeding on reproductive ability in a noncompetitive environment were found to be minor by comparison. The maintenance of captive populations under noncompetitive conditions can therefore be expected to minimize adaptive changes due to natural selection in the changed environment.     

Inactivation of coliphage MS-2 and poliovirus by copper, silver, and chlorine.

The efficacy of electrolytically generated copper and silver ions (400 and 40 micrograms/L, respectively) was evaluated separately and in combination with free chlorine (0.2 and 0.3 mg/L) for the inactivation of coliphage MS-2 and poliovirus type 1 in water at pH 7.3. The inactivation rate was calculated as log10 reduction/min: k = -(log10 Ct/C0)/t. The inactivation of both viruses was at least 100 times slower in water containing 400 and 40 micrograms/L copper and silver, respectively (k = 0.023 and 0.0006 for MS-2 and poliovirus, respectively), compared with water containing 0.3 mg/L free chlorine (k = 4.88 and 0.036). Significant increases in the inactivation rates of both viruses were observed in test systems containing 400 and 40 micrograms/L copper and silver, respectively, with 0.3 mg/L free chlorine when compared with the water systems containing either metals or free chlorine alone. Poliovirus was approximately 10 times more resistant to the disinfectants than coliphage MS-2. This observation suggests either a synergistic or an additive effect between the metals and chlorine for inactivation of enteric viruses. Use of copper and silver ions in water systems currently used in swimming pools and spas may provide an alternative to high levels of chlorination.     

Endogenic development of sediments in a eutrophic lake

Sedimentation in the Gouet reservoir (France), measured for 2 years at 8 stations, was maximal during summer, when river inputs were minimal. Physical and chemical conditions in the deposits indicate that the endogenous part of sedimentation was about 70% and resulted from significant diatom production. The high sedimentation rate on the bottom was favoured by the funnel morphology of the reservoir, the chronic lack of oxygen in the water column, and the repeated copper sulfate treatment. The former river meanders of the reservoir were the preferential deposit sites.     

Variation of the oviposition preferences of Aedes aegypti in function of substratum and humidity

Two Aedes aegypti (L.) populations were studied in the laboratory regarding the preference for three types of breeding sites, i.e., flasks containing only water, flasks with a plant and flasks with a stick. Each of these breeding units was placed in one cage and the choice of the oviposition sites was determined for individual females and three females per experimental unit at two humidity levels. Preference for ovipositing on the water surface was observed and varied according to experimental unit and humidity. Mean hatching of eggs in water surface was 46.6%. Experiments with three females showed a more marked difference than when only one female was used. Inter and intrapopulation variability regarding oviposition sites was observed. The discrimination between the different oviposition substrates, hatching in water surface and its implication for mosquito control are discussed.     

Aedes albopictus in Rome: results and perspectives after 10 years of monitoring

In 1997, Aedes albopictus (Skuse 1894) was detected in Rome in two opposite areas of the city. In the following 2 years, the species quickly spread. In 2000, scattered foci of the species were reported in the whole urban area and in the outskirts of the capital city. In Rome, Ae. albopictus seems to have found optimal environmental conditions to proliferate and to overwinter through and without diapausing eggs. In ten years Ae. albopictus has colonized the whole urban area through three phases: first massive spread, following maintenance of infestation, and colonization of alternative winter breeding sites with favorable climatic conditions. Data collected during the 2007 show that rainfall is no longer the most important factor for the development of the species, with respect to the past. In fact Ae. albopictus probably has found new alternative larval breeding sites through the colonization of small water collections refilled periodically by human activities. During 2007-2008 winter season, in order to evaluate the species adaptability, a study of eggs hatching and length of larval cycle at low temperatures, was carried out in laboratory and in simulated field conditions. Data and results are showed and discussed also by the light of existing literature.     

Kinetic and spectroscopic studies of the interaction of copper with dopamine beta-hydroxylase

The question of the stoichiometry of copper bound to dopamine beta-hydroxylase and the number of copper atoms required for maximal activity was addressed in this study. Incubation of tetrameric enzyme from bovine adrenal medulla with 64Cu2+ followed by rapid gel filtration yielded an enzyme containing 8.3-8.9 mol of Cu/mol of tetramer. An identical stoichiometry was obtained by analysis of bound copper by atomic absorption methods. NMR and EPR were used to monitor titrations of the enzyme with Cu2+ and showed that the longitudinal relaxation rate of solvent water protons and the amplitude of the signal at g approximately 2 increased linearly up to a copper to protein ratio of approximately 8. Additional titrations also indicate that an enzyme-Cu2+-tyramine-CN- inhibitory complex was formed when 8 mol of Cu2+ are bound per mol of enzyme. The rate of inactivation of dopamine beta-hydroxylase by the mechanism-based inhibitor 2-Br-3-(p-hydroxyphenyl)-1-propene was measured and used as a method to follow enzymatic catalysis. An increase in rate was observed with increasing Cu2+ up to a protein to Cu2+ ratio of 8 Cu/tetramer. The rate becomes constant after this ratio is achieved. These data indicate that dopamine beta-hydroxylase specifically binds 8 mol of Cu/tetramer and that this stoichiometry is required for maximal activity.     

The naturally derived insecticide spinosad is highly toxic to Aedes and Anopheles mosquito larvae

Spinosad is a naturally derived biorational insecticide with an environmentally favourable toxicity profile, so we investigated its potency against mosquito larvae (Diptera: Culicidae). By laboratory bioassays of a suspension concentrate formulation of spinosad (Tracer), the 24 h lethal concentration (LC50) against Aedes aegypti (L.) third and fourth instars was estimated at 0.025 p.p.m. following logit regression. The concentration-mortality response of third- and fourth-instar Anopheles albimanus Weidemann did not conform to a logit model. The LC50 value of spinosad in Anopheles albimanus was 0.024 p.p.m. by quadratic linear regression. A field trial in southern Mexico demonstrated that spinosad 1 p.p.m. compared with the standard temephos (Abate) 1% granules 100 g/m3 water prevented Ae. aegypti breeding in plastic containers of water for 8 weeks; at 10 p.p.m. spinosad prevented breeding for > 22 weeks. In another field trial, spinosad at 5 p.p.m. and temephos both completely eliminated reproduction of Ae. aegypti for 13 weeks. In contrast, the bacterial insecticide Bacillus thuringiensis var. israelensis (Bti, Vectobac) AS) performed poorly with just 2 weeks of complete inhibition of Ae. aegypti breeding. Spinosad also effectively prevented breeding of Culex mosquitoes and chironomids in both trials to a degree similar to that of temephos. We conclude that spinosad merits evaluation as a replacement for organophosphate or Bti treatment of domestic water tanks in Mesoamerica. We also predict that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites.     

A transport kinetic concept for ion uptake by plants

Summary A previously presented transport kinetic model for ion uptake by plants was tested by using data for copper uptake by barley grown in soil or water culture and corresponding data for copper concentration in the soil solution or culture solution. As no significant differences were obtained between experimentally determined values for copper uptake and those calculated by using the model, it was concluded that the experimental evidence was consistent with the proposed transport kinetic model. The model allows the Michaelis-Menten Constant of ion uptake and the Mean Maximal Rate of ion uptake to be calculated for various time intervals of the growth period of plants. The Michaelis-Menten Constant for copper uptake by barley grown in soil was the same as that for plants grown in water culture, provided that the soil solution and the water culture solution had the same chemical composition. The concentration of copper in the soil solution, at which the rate of copper uptake was zero, was 2–4 times larger than corresponding values obtained in water culture solution. re]19751028     

Determination of the taste threshold of copper in water

Copper effects on human health represent a relevant issue in modern nutrition. One of the difficulties in assessing the early, acute effects of copper ingested via drinking water is that the taste of copper may influence the response and the capacity to taste copper in different waters is unknown. The purpose of the study was to determine the taste threshold of copper in different types of water, using soluble and insoluble salts (copper sulfate and copper chloride). Copper-containing solutions (range 1.0-8.0 mg/l Cu) were prepared in tap water, distilled deionized water and uncarbonated mineral water. Sixty-one healthy volunteers (17-50 years of age), with no previous training for sensory evaluation, participated in the study. A modified triangle test was used to define the taste threshold value. The threshold was defined as the lowest copper concentration detected by 50% of the subjects assessed. To evaluate the olfactory input in the threshold value obtained, 15 of 61 subjects underwent a second set of triangle tests with the nose open and clamped, using distilled water with copper sulfate at a concentration corresponding to the individual's threshold. The taste threshold in tap water was 2.6 mg/l Cu for both copper sulfate and copper chloride. The corresponding values for distilled deionized water were 2.4 and 2.5 mg/l Cu for copper sulfate and copper chloride, respectively. In uncarbonated mineral water the threshold values were slightly higher, 3.5 and 3.8 mg/l Cu for copper sulfate and for copper chloride, respectively, which are significantly higher than those observed in tap and distilled waters (P < 0.01, Kruskal-Wallis test). The taste threshold did not change significantly when the nose was clamped. In conclusion, the median values for copper taste threshold were low, ranging between 2.4 and 3.8 mg/l Cu, depending on the type of water.     

Oxygen concentration profiles and exchange in sediment cores with circulated overlying water

SUMMARY. 1. The overlying water of intact sediment cores was constantly stirred with an impeller at a rate sufficient to mix turbulently the water column and maintain the diffusive boundary layer at a determined thickness. The system allowed standardization of water circulation in laboratory sediment core experiments.
2. Both oxygen concentration and oxygen penetration depth in the sediments decreased, the former by 70% and the latter from 4.2 mm to 2.0 mm, when the overlying water was not stirred for 24 h, as measured with oxygen microelectrodes in a lake sediment core.
3. Oxygen profiles measured in sediment cores in the laboratory were similar to those measured in situ when the overlying water was stirred with an impeller at such a rate that a similar thickness of the diffusive boundary layer at the sediment-water interface developed in the laboratory as that in situ.
4. Sediment oxygen consumption was calculated from: (1) measured oxygen profiles in the diffusive boundary layer and the molecular diffusion coefficient for oxygen in water; (2) the measured oxygen decrease in the top of the sediments and the estimated diffusion coefficient in the sediment; and (3) by oxygen differences in the overlying water after incubation of sediment cores.