Accumulation and Elimination of Coliphage S-13 by the Hard Clam, Mercenaria mercenaria

Accumulation and elimination of viral particles by hard clams, Mercenaria mercenaria, were studied with the coliphage S-13 as a working model. Escherichia coli uptake and elimination were simultaneously monitored. Clams were exposed to low levels of S-13 (7 particles/ml) in running seawater for several days, achieving titers in tissues from 2 to more than 1,000 times the levels to which they had been exposed. Bacterial accumulation (previously established by other workers) was comparable. Upon exposure to virus-free running water, clams polluted to relatively low levels (100 plaque-forming units/ml) eliminated most of their bacterial contaminants in 24 to 48 hr. Viral contaminants, however, persisted for several days to weeks even under ideal conditions for clam activity, provided that the temperature remained below the inactivation threshold for the virus. Most of the accumulated virus appeared to be sequestered in the digestive gland. These sequestered particles are refractory to those mechanisms responsible for elimination of bacterial contaminants. This discrepancy points out the need for caution in evaluating the efficiency of shellfish depuration processes, especially if only a bacterial criterion is used as a monitoring system.     

Vivo clearance of enteric bacteria from the hemolymph of the hard clam and the American oyster.

American oysters, Crassostrea virginica, and hard clams, Mercenaria mercenaria, were experimentally contaminated with Escherichia coli, Salmonella typhimurium, and Shigella flexneri either by intracardial injection or via the natural route of ingestion. Bacterial inactivation in the hemolymph was monitored for 72 h after exposure to these enteric pathogens at 20 and 6 degrees C. At 6 degrees C, both mean bacterial uptake by ingestion and subsequent clearance was singificantly lower that at 20 degrees C. However, substantial bacterial clearance from the hemolymph occurred for both shellfish at each temperature. At 20 degrees C, viable bacteria were no longer detectable after 24 h in hemolymph of either clams or oysters after exposure to contaminated water containing 4 x 10(3) bacteria per ml.     

Vivo clearance of enteric bacteria from the hemolymph of the hard clam and the American oyster.

American oysters, Crassostrea virginica, and hard clams, Mercenaria mercenaria, were experimentally contaminated with Escherichia coli, Salmonella typhimurium, and Shigella flexneri either by intracardial injection or via the natural route of ingestion. Bacterial inactivation in the hemolymph was monitored for 72 h after exposure to these enteric pathogens at 20 and 6 degrees C. At 6 degrees C, both mean bacterial uptake by ingestion and subsequent clearance was singificantly lower that at 20 degrees C. However, substantial bacterial clearance from the hemolymph occurred for both shellfish at each temperature. At 20 degrees C, viable bacteria were no longer detectable after 24 h in hemolymph of either clams or oysters after exposure to contaminated water containing 4 x 10(3) bacteria per ml.     

Pre-treating paclobutrazol enhanced chilling tolerance of sweetpotato

The objective of this work was to study changes in low molecular weight antioxidants and antioxidative enzymes in chilling-stressed sweetpotato, as affected by paclobutrazol (PBZ) pre-treatment 24 h prior to exposure to chilling conditions. Sweetpotato ‘TN71’ and ‘TN65’ were treated with 300 mg PBZ/5 ml/plant, after which plants were subjected to 7°C/7°C (day/night) for periods of 1, 3 and 5 days, followed by a 3-day recovery period at 24°C/20°C (day/night). A factorial experiment in completely randomized design with four replications was used in this study. Young fully expanded leaves at each temperature and period of time were clipped for antioxidative system measurement. We concluded that different varieties displayed variations in their oxidative system, and the differential expressions of each genotype were associated with chilling stress response. Plants with various antioxidative systems responded differently to chilling stress according to the duration of the chilling period and subsequent re-warming period. ASA, GSH and GSSG contents were enhanced in TN71 prior to chilling stress. Increased APX, GR, ASA and MDA activities accounted for chilling tolerance in TN65. Furthermore, our results indicate that the elevated levels of the antioxidative system observed after PBZ pre-treatments afforded the sweetpotato leaf improved chilling-stress tolerance. The levels of ASA and GSSG of both TN71 and TN65 under chilling were significantly raised by pre-treating with PBZ. PBZ pre-treatment exhibited the important function of enhancing the restoration of leaf oxidative damage under chilling stress and increasing the chilling tolerance of plants to mitigate chilling stress effects.     

重金属Cd胁迫对红树蚬的抗氧化酶、消化酶活性和MDA含量的影响

摘要：本文在实验室条件下,观测不同Cd胁迫处理（时间和水平）对红树蚬（Geloina coaxans (Gmelin)）的胃组织中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性、丙二醛(MDA)含量,及淀粉酶、脂肪酶和蛋白酶等3种消化酶活性的影响效应。结果表明,高浓度组（4.00 mg?L-1和8.00 mg?L-1）在较短的暴露时间即出现SOD 活性显著诱导,而低浓度组则需要更长的暴露时间。所有处理组的CAT活性均先受诱导而后抑制,但去除胁迫后低浓度组活性上升,高浓度组则持续下降。低浓度处理组在胁迫初期MDA含量上升,但随后下降至较低水平;高浓度处理组MDA含量上升稍为滞后,但随后上升至较高水平。在消化酶方面,Cd对红树蚬胃组织淀粉酶的影响基本上表现为抑制;在胁迫初期脂肪酶活性受到显著诱导,随着胁迫时间延长酶活性则下降,同时胁迫解除后不同程度恢复;对蛋白酶的影响效应规律性不明显。显著的效应-剂量间相关关系存在于特定时间的SOD活性（1d）,CAT、淀粉酶活性和MDA含量（7d和恢复6d）。本文还探讨了这些指标作为生物标记物应用于监测海洋重金属污染的可能性。 关键词：红树林;红树蚬;Cd胁迫;消化酶;抗氧化酶;脂质过氧化     

Pathology of acute poisoning with the insecticide Sevin in the bent-nosed clam, Macoma nasuta

Toxicity tests of 96-hr duration of the insecticide Sevin were done with adult bent-nosed clams, Macoma nasuta. Sevin concentrations of 15, 20, 25, and 30 mg/liter were used in duplicate tests. The criterion of “death” was the inability of clams to retract siphons or to close valves. About half of the animals so affected were removed from the test solutions and returned to clean sea water to observe if recovery occurred, and others were preserved for histological examination.No “dead” clams recovered within 96 hr after return to clean water. The histopathology consisted primarily of necrosis of epithelial tissue of the gill, mantle, siphon, and suprabranchial gland, and the severity of damage was directly related to the test concentrations. Vacuolization, rupture, and pycnosis of cells occurred. The gills were the most severely affected organs. Epithelial cells of the gill filaments bearing the frontal, laterofrontal, and lateral cilia were sloughed as early as 24 hr after the beginning of exposure to Sevin. About 50% of the exposed clams had lost one or both siphons and also the epithelia on still attached segments within 96 hr of exposure. There were no deaths of control clams, and their tissues were normal.     

Modulation of the antioxidative response of Spartina densiflora against iron exposure

Spartina densiflora , an invader cordgrass living in polluted salt marshes of the Odiel estuary (SW Spain), was collected and cultured under controlled laboratory conditions. After acclimation to non-polluted soils for 28 days, both metabolites and enzymes activities used as indicators of oxidative stress were reduced significantly. Then, plants were exposed to 500 and 1000 ppm Fe-ethylenediamine-N,N'-2-hydroxyphenyl acetic acid (EDDHA) for 28 days. Our data demonstrate that iron content in leaves was enhanced by iron exposure. This iron increase caused an enhancement in the concentration of H₂O₂, hydroperoxides and lipid peroxidation, and a decrease in chlorophyll levels. Thus, iron exposure led to oxidative stress conditions. However, oxidative indicators stabilised after first 2 weeks of exposure, although the highest iron levels in leaves were reached at the end of treatments. Iron exposure induced an enhancement of catalase, ascorbate peroxidase and guaiacol peroxidase activities, together with an increase in total and oxidised ascorbate. This response may be defensive against oxidative stress and thus help to explain why cell oxidative damages were stabilised. Thus, by using a sensitive long-time protocol, iron-dependent oxidative damages may be controlled and even reverted successfully by the activation of the antioxidative defences of S. densiflora . This efficient antioxidative system, rapidly modulated in response to excess iron and other environmental stressors, may account for S. densiflora 's successful adaptation to stress conditions in its habitat.     

Effects of Prolonged Chlorine Exposures upon PCR Detection of Helicobacter pylori DNA

The effect of low doses of free chlorine on the detection of Helicobacter pylori (H. pylori) cells by qPCR in tap water was monitored. Detection of sequences targeted to the ureA gene from preparations containing 107 cells/ml decreased about 2-4 logs by days 9 and 14, respectively. When duplicate suspensions of the 107 cells/ml were exposed to higher levels of chlorine, 0.2-2.2 mg/l, by day 9 and 14 there were 5 and 6 log decreases, respectively, in the detection of ureA gene. H. pylori target sequences (within suspended, intact cells at densities of 102-103 cells /ml) were rendered undetectable by qPCR analysis after 17 h of continuous exposure to low chlorine levels common to treated drinking water distribution systems. The persistence of DNA sequences within treated distribution systems detectable by qPCR may be as brief as 17 h especially for bacteria such as H. pylori which are known to occur in very low numbers within treated distribution systems. This study suggests that degradation of H. pylori DNA target sequences by chlorine levels commonly found within treated water distribution systems occurs within the average water retention times (2-3 days) commonly found in these systems.     

Environmental Cadmium Exposure Impacts Physiological Responses in Manila clams

The physiological responses of marine bivalves to chronic cadmium (Cd) exposure at sub-lethal concentrations have been well documented. As of now, few studies have examined the effect of Cd exposure and subsequent recovery period at environmentally realistic concentrations. In this study, environmentally, Cd exposures were performed to assess the physiological responses of the Manila clam Ruditapes philippinarum. The clams were exposed to waterborne Cd at two environmentally realistic concentrations (4 and 40 μg L^?1) for 35 days and then allowed to recover for another 35 days. The accumulation and elimination of Cd in R. philippinarum were tissue-specific and dose- and time-dependent. Cd accumulation increased sharply in the digestive gland, and Cd elimination was rapid in the gill. Major physiological responses, including clearance rate, absorption efficiency, respiration rate, excretion rate, oxygen to nitrogen ratio, and scope for growth, were significantly affected by Cd exposure. Yet, the clams exposed to 4-μg L^?1 Cd were able to quickly recover their normal physiological processes and clearly exhibited catch-up growth once they were transferred to clean seawater. Hence, R. philippinarum can exhibit good physiological plasticity when confronted with moderately environmental Cd exposure. All physiological responses measured exhibited a highly significant and generally predictable correlation with tissue Cd concentration, which in turn, reflected environmentally realistic exposure conditions. Our results further confirm that the measurement of physiological responses is a sensitive method for assessing stress at environmentally realistic metal concentrations.     

Paclobutrazol pre-treatment enhanced flooding tolerance of sweet potato

The objective of this experiment was to study changes of antioxidants and antioxidative enzymes in the flooding-stressed sweet potato leaf, as affected by paclobutrazol (PBZ) treatment at 24 h prior to flooding. Sweet potato 'Taoyuan 2' were treated with 0 and 0.5 mg/plant of PBZ, afterwards subjected to non-flooding and flooding-stress conditions for 0, 1, 3, and 5 d, followed by a 2 d drainage period. The study was conducted as a factorial experiment in completely randomized blocks with three replications maintained within a screen house. Plants with various antioxidative systems responded differently to flooding stress according to the duration of the flooding period and subsequent drainage period. The increased levels of antioxidants and antioxidative enzymes observed on different days of flooding afforded the sweet potato leaf with improved flooding tolerance. Glutathione reductase activity in the leaf was significantly enhanced over 5 d continuous flooding followed by a drainage period, in comparison with non-flooding conditions. Under non-flooding conditions, antioxidative system of leaf was regulated and elevated by PBZ pre-treatment. PBZ treatment may enable sweet potato 'Taoyuan 2' to maintain the balance between the formation and the detoxification of activated oxygen species. Our results also show that under flooding-stress conditions, the level of 'Taoyuan 2' antioxidative system is linked to PBZ treatment. Pre-treating with PBZ may increase levels of various components of antioxidative systems after exposure to different durations of flooding and drainage, thus inducing flooding tolerance. PBZ exhibited the important function of enhancing the restoration of leaf oxidative damage under flooding stress after the pre-application of 0.5 mg/plant. These findings may have greater significance for farming in frequently flooded areas.     

Effects of Substerilization Doses of Co60 Gamma Radiation on the Cold-Storage Life Extension of Shucked Soft-Shelled Clams and Haddock Fillets

Total aerobic-facultative and anaerobic (clostridia) macrocolony count data are presented, with analysis and interpretation, for both haddock fillets and shucked soft-shelled clams which received doses of from 50,000 to 800,000 rad of Co⁶⁰ gamma rays. These data indicated that haddock fillets may be maintained in good condition at refrigeration temperatures above freezing for about 1 week at 6 C, and approximately 2 weeks at 0 C, when treated with from 50,000 to 150,000 rad of ionizing radiation. In the dose range from 200,000 to 350,000 rad, the storage life may be extended up to some 2 weeks at 6 C, and 3 weeks at 0 C. Treatments in the dose range from 400,000 to 500,000 rad may defer spoilage for about 1 month, and doses of 550,000 to 650,000 rad afford protection against bacterial spoilage up to approximately 1.5 months. At the high substerilization doses of 700,000 to 800,000 rad, haddock fillets may be held for from 2 to 3 months in refrigerated storage before becoming unfit for marketing and consumption. Shucked soft-shelled clams can be held for about 2.5 weeks at 0 C and close to 12 days at 6 C, when given low substerilization doses of from 50,000 to 150,000 rad of ionizing radiation. At doses of from 200,000 to 350,000 rad, the clams may be preserved effectively for periods up to 3 weeks at 0 or 6 C, and some 6 weeks at these temperatures with doses of about 450,000 rad. With treatments of 500,000 to 600,000 rad, the storage life may be extended for some 2 months, and at doses of 650,000 to 800,000 rad the shucked clams remain in a good state of preservation for up to 3 months at temperatures of 0 to 6 C. Thus, it would appear that shucked soft-shelled clams may be maintained for significantly longer periods in refrigerated storage than haddock fillets when the same radiation treatments are applied to each product. Clostridia levels in both products were relatively low initially, and were reduced significantly by the gamma rays at the doses studied. Moreover, those clostridia that survived the radiation treatments were found to remain at safe, low levels during the various periods in refrigerated storage employed for these products, a very encouraging result from the public health, as well as commercial, standpoint.     

A Preliminary Laboratory Investigation of PCB Flux from Dredge Resuspensions and Residuals

A preliminary laboratory investigation was conducted to understand the relative contributions of major dredge resuspension and residual processes on the releases of polychlorinated biphenyl (PCB) contaminants from sediments to water column. Sediments from New Bedford Harbor were used as test samples. Six sets of experiments were run for simulated resuspension and residual scenarios. During the experiments, water above the sediments was recirculated by peristaltic pumping or orbital shaking and the levels of two PCBs, Aroclor 1248 (PCB-1248) and Aroclor 1254 (PCB-1254), were monitored for 15 days. Analysis of the model predicted data indicated that resulting water column PCB concentrations differed with sediment surface, residual, and resuspension type. Highest PCB water column concentrations were observed for a condition which used a settled fluff from thin sediment slurry as a residual source and the column water was recirculated by orbital shaking. Lowest water column PCB levels were observed for a thick sediment deposit placed over clean sand. The PCB levels in the water column for all six simulated conditions were several orders higher than the USEPA ambient water quality criteria concentrations for aquatic environment and human consumption.     

Changes in hypothalamic noradrenaline and 5-hydroxytryptamine levels during the development of warm- and cold-adaptation in the rat

Hypothalamic 5-hydroxytryptamine (5-HT) and noradrenaline (NA) as well as plasma corticosterone levels were studied in male rats after 1, 2, 4 and 6 weeks of exposure to 4--7 or 30--31 degrees C. An increase of the NA concentration and a decrease of the 5-HT level was observed after the first week in both cold and warm environment together with an increase of plasma corticosterone levels in both groups. NA, 5-HT and plasma corticosterone levels returned to normal in cold-exposed animals by the 6th week whereas in warm-acclimated rats NA and corticosterone levels regained their initial values and 5-HT concentrations remained low. Changes by the end of the first week of exposure may result from the thermal stress. The low 5-HT levels of warm-adapted animals at the end of the 6th week were probably secondary to the process of adaptation.     

At the tipping point: Differential influences of warming and deoxygenation on the survival,emergence, and respiration of cosmopolitan clams

Although warming and low dissolved oxygen (DO) levels are co‐occurring significant climatic stressors in the ocean, the combined effects of these stressors on marine benthic animals have not been well established. Here, we tested the effects of elevated temperatures and low dissolved oxygen levels on the survival, emerging behavior from sediment, and the respiration of juvenile cosmopolitan Manila clams (Venerupis philippinarum) by exposing them to two temperatures (20 and 23.5°C) and DO levels (3.5 and 6–7 mg/L). Although within previously described tolerable ranges of temperature and DO, this 3.5°C increase in temperature combined with a 50% decrease in DO had a devastating effect on the survival of clams (85% mortality after 8 days). The mortality of clams under normoxia at 23.5°C appeared to be higher than under the low DO condition at 20°C. On the other hand, more clams emerged from sediment under the low DO condition at 20°C than under any other conditions. Oxygen consumption rates were not significantly affected by different conditions. Our results suggest temperature elevation combined with low oxygen additively increases stress on Manila clams and that warming is at least as stressful as low DO in terms of mortality. However, low DO poses another threat as it may induce emergence from sediment, and, thus increase predation risk. This is the first evidence that a combination of warming and deoxygenation stressors should reduce population survival of clams much more so than changes in a single stressor.     

Oxidative stress,growth factor production and budding in potato tubers during cold storage

In order to verify the role played by oxidation in the budding of potato tubers (Solanum tuberosum L. cv. Kennebec), the physiological events occurring below bud at 4°C have been studied for a period of 6 months. The low temperature storage induced an increase in the degree of unsaturation and a decrease in the ratio of saturated/unsaturated fatty acids of membrane polar lipids with a subsequent increase of lipid hydroperoxides (LOOH). Cold stress increased both enzymatic antioxidative activities (superoxide dismutase, SOD, E.C.1.15.1.1; catalase, CAT, E.C. 1.11.1.6), and α-tocopherol levels thus protecting membrane's polyunsaturated lipids. Between 0 and 15 days of storage SOD/CAT ratio, α-tocopherol, LOOH levels and the degree of lipid unsaturation showed strong variations. After 30 to 120/150 days the antioxidative system seemed to reach a homeostasis different from that of time 0, accompanied by a constant increase of indole-3-acetic acid (IAA) after 60 days. The antioxidative system, after 150 days, lost its efficiency while LOOH levels were maintained higher than time 0 and IAA concentration was sufficient to allow sprouting.     

Oxidative stress, growth factor production and budding in potato tubers during cold storage.

In order to verify the role played by oxidation in the budding of potato tubers (Solanum tuberosum L. cv. Kennebec), the physiological events occurring below bud at 4 degrees C have been studied for a period of 6 months. The low temperature storage induced an increase in the degree of unsaturation and a decrease in the ratio of saturated/unsaturated fatty acids of membrane polar lipids with a subsequent increase of lipid hydroperoxides (LOOH). Cold stress increased both enzymatic antioxidative activities (superoxide dismutase, SOD, E.C.1.15.1.1; catalase, CAT, E.C.1.11.1.6), and alpha-tocopherol levels thus protecting membrane's polyunsaturated lipids. Between 0 and 15 days of storage SOD/CAT ratio, alpha-tocopherol, LOOH levels and the degree of lipid unsaturation showed strong variations. After 30 to 120/150 days the antioxidative system seemed to reach a homeostasis different from that of time 0, accompanied by a constant increase of indole-3-acetic acid (IAA) after 60 days. The antioxidative system, after 150 days, lost its efficiency while LOOH levels were maintained higher than time 0 and IAA concentration was sufficient to allow sprouting.     

Proline accumulation and the adaptation of cultured plant cells to water stress

The transfer of cultured tomato cells (Lycopersicon esculentum cv VFNT-Cherry) to a low water potential environment resulted in an increased dry weight to fresh weight ratio accompanied by a rapid accumulation of proline. Proline content continued to increase as osmotic adjustment and growth occurred. The initial increase in proline concentration was accompanied by a drop in turgor. However, proline levels continued to increase with a gain in turgor during osmotic adjustment. Thus, the accumulation of proline depended not only on cell water potential, or on the initial loss of turgor but more closely on cell osmotic potential. The ultimate level of proline depended on the level of adaptation. Proline levels remained high after more than 100 cell generations in low water potential media, but declined rapidly after transfer to media with a less negative water potential. Addition of exogenous proline to the medium during water stress and during osmotic downshock alleviated the normally resulting inhibition of growth. The results suggest a positive role for proline accumulation in adaptation of cells to changing external water potentials.     

雌二醇、壬基酚、多氯联苯、镉和锌暴露对唐鱼体内超氧化物歧化酶活性的影响

目的 研究雌二醇(E2)、壬基酚(NP)、多氯联苯(PCBs)、镉(Cd2+)和锌(Zn2+)单独以及联合暴露对唐鱼体内SOD酶活力的影响.方法设计不同浓度的单一物质及混合物对唐鱼14 d暴露染毒,定量测定7、14d体内SOD酶活力的变化.结果 ①低浓度E2处理唐鱼7d可诱导SOD活性显著上升,时间延长、浓度升高时SOD活性无明显变化;②低浓度NP对SOD活性没有明显的影响,高浓度NP使SOD活性极显著上升;③中高浓度组PCBs处理7、14 d,低浓度组PCBs处理14 d时SOD活性被抑制,抑制程度随着时间的延长和浓度的升高有增强的趋势;④Cd2+、Zn2+各浓度组都对唐鱼体内的SOD活性产生了一定的抑制作用,并且抑制程度随着时间的延长而加深.结论 E2、NP、PCBs、Cd2+、Zn2+对唐鱼的SOD酶活力有明显影响,它们单独作用时的SOD活性与暴露浓度之间存在良好的剂量-效应关系.联合作用效应与暴露时间和(或)各物质浓度有关,大部分表现为毒性增强.     

Aluminium accumulation and distribution in the freshwater clams (unionidae)

1. Freshwater unionids (Anodonta anatina L. and Unio pictorum L.) were exposed to aluminium (300 and 900 μg/l, nominal) in continuous (3 weeks) and fluctuating (24 days) acid exposures.2. In addition, accumulation was monitored for 2 weeks under semi-static acid (pH4-5) and circumneutral (pH 6.6-8.3) conditions in hard (35 mg Ca/l) and soft water (3.5 mg Ca/l).3. In addition, a fluctuating exposure of 24 days, consisting of three intermittent pulses which combined low pH (4–5) and high Al (900 μg/l) concentration, was performed.4. The various organs of A. cygnea, collected from a watershed with relatively high heavy metal concentrations, were analyzed for their Al and Cd concentrations.5. The ultimate order of the Al and Cd concentration in these clams was identical: kidney ≥ midgut gland ≥ rest ≥ gill ≥ mantle.6. During the 3 weeks of exposure, the Al concentration in the gills and kidney increased linearly, and saturation level was not reached.7. The Al concentration in the calcium concretion material isolated from the gills was lower than that of the whole organ.8. An elimination period of 12 weeks was needed to reach the background level of Al in the gills, whereas in the kidney the initial Al concentration was reached after 4 weeks of elimination.9. In both species, the ambient pH had a significant effect on the Al accumulation in the gills, whereas the effect of the water hardness was only of minor importance.10. Rapid elimination of the Al accumulated in the gills of U. pictorum during the episodic 3-day exposure was recorded.