Factors influencing the attachment strength of Dreissena polymorpha (Bivalvia)

Abstract

The effects of several factors (shell length, exposure time, substratum orientation in space, illumination, temperature, conspecifics) upon the attachment strength (measured with a digital dynamometer) of the freshwater, gregarious bivalve Dreissena polymorpha were studied under laboratory conditions. A rapid increase in attachment strength was observed on resocart (a thermosetting polymer based on phenol-formaldehyde resin, with paper as filler) substrata during the first 4-d exposure, after which it stabilised at ca 1 N. The attachment strength increased also with mussel size. Mussel adhesion on variously oriented surfaces (vertical, upper horizontal and lower horizontal) was similar. Illumination inhibited attachment strength, as expected for a photophobic species, but only after a 2-d exposure. After 6 d, no effects of light were detected. Thus, illumination seemed to influence the attachment rate, rather than the final strength. The optimum temperature for mussel attachment was 20 – 25°C. At lower and higher temperatures (5 – 15°C and 30°C), their adhesion strength decreased. The presence of conspecifics stimulated mussel attachment strength.     

Quantitative studies of increased cell-to-substratum adhesion at low temperature

The cell-to-substratum adhesion of an established epithelial cell line cultured for 24 h on glass coverslips was determined at 4 degrees C, 8 degrees C and 37 degrees C using a miniaturised parallel-plate shearing apparatus. The measurements of the minimum shear necessary to dislodge the cells (minimum distraction force, MDF) demonstrated a three- to fourfold increase in the adhesion of the cells at 4 degrees C (6.17 Pa) compared to that at 37 degrees C (1.36 Pa). At 8 degrees C the MDF was 2.31 pascals. Part of the adhesion was resistant to mild trypsinisation. Trypsin-resistant adhesion (TRA) was stabilised by low temperature, and by treatment with concanavalin A (50 micrograms ml-1) or colchicine (200-400 microM). The effects of con A (140 micrograms ml-1) and low temperature (4 degrees C) were additive, giving a combined MDF of greater than 9.27 pascals. On the basis of their different temperature and protease susceptibility it is suggested that trypsin-sensitive adhesion (TSA) and TRA represent separate functional classes of cell-to-substratum attachment corresponding to 'frictional' and 'tractional' adhesion, respectively.     

Does rock type account for variation in mussel attachment strength? A test with <Emphasis Type="Italic">Brachidontes rodriguezii</Emphasis> in the southwestern Atlantic

Mussel attachment strength varies in space and time, frequently in association with variations in wave exposure. Yet, it remains uninvestigated whether different rock types can contribute to variation in mussel attachment. Here we compared the attachment strength of the mussel Brachidontes rodriguezii between soft and hard intertidal rock substrates that are typical of coastal Buenos Aires Province, Argentina: Pampean loess cemented by calcium carbonate and orthoquartzite, respectively. Overall comparisons of mussel attachment across natural platforms of either rock type (10 loess sites and 4 orthoquartzite sites) indicated stronger mussel attachment to orthoquartzite. However, mussel attachment strength did not differ when compared across natural loess platforms and introduced orthoquartzite blocks (i.e., groins and revetments) occurring within the same site. Mussels attaching to loess showed more byssal threads than those attaching to orthoquartzite at the same site. These findings suggest, first, that rock type does not influence mussel attachment strength in our study system, secondly, that overall differences in mussel attachment strength with rock type across natural platforms in our study range are due to confounding influences of co-varying factors (e.g., wave exposure) and, finally, that mussels can increase byssus production to counteract potential substrate failure when attaching to soft, friable rock. The latter likely explains the ability of mussels to maintain relatively stable cover across rocks of contrasting hardness.     

Influence of light intensities on dressing behavior in elderly people

With civilized environments in modern society, since the people tend to depend more on artificial illumination than on natural illumination which makes less discrepancy between day and night life, clarifying the relationship between human life and illumination is necessary. In our previous studies, we found that the subjects dressed faster with thicker clothing in the morning than in the evening when the room temperature decreased from 30 degrees C to 15 degrees C over 1 hour. We considered these results in terms of load error between the actual and set-point values in the core temperature. The present study was designed to examine the effect of bright light (3,000 lx)/dim light (50 lx) exposure (09:30 h-14:30 h) on dressing behavior and thermoregulatory responses in the elderly people during the afternoon cold exposure. Five female subjects were instructed to dress to feel comfortable when the room temperature was decreased from 30 degrees C to 15 degrees C (15:00 h-17:00 h). The subjects felt cooler and dressed more quickly with thicker clothing after dim light exposure, it is conceivable that the set-point value of core temperature is reduced under the bright light condition. We discussed these results in terms of the establishment of set-point values in the core temperature at bright light condition. If the set-point of the core temperature is lower in the bright than in the dim light condition in present experiment, the dressing behavior with thinner clothing in the bright light condition is advantageous, since it enables the core temperature to reach its set-point value more easily.     

Teratogenicity of 27.12-MHz radiation in rats is related to duration of hyperthermic exposure

Five groups of pregnant Sprague-Dawley rats were either sham exposed or were irradiated in a 27.12-MHz radiofrequency (RF) field at 55 A/m and 300 V/m on gestation day 9. The absorbed power (approximately 11 W/kg) caused a relatively rapid increase in the rat's colonic temperature. Rats in group I were sham irradiated for 2.5 h at 0 A/m, 0 V/m. In group II RF irradiation was terminated after the rat's colonic temperature reached 41.0 degrees C. In group III the 41.0- degrees C temperature was maintained an additional 2 h by manually varying the incident field strength. In group IV irradiation was terminated after the rat's colonic temperature reached 42.0 degrees C. In group V the 42.0- degrees C temperature was maintained an additional 15 min by varying the field strength. At both temperatures the teratogenic and embryotoxic effects of the RF-induced hyperthermia increased as the exposure duration increased, but the increase was especially noticeable at 42.0 degrees C. The results indicate that the teratogenic and embryotoxic effects of RF-induced hyperthermia are related to both the temperature of the dam during exposure and the length of time the dam's temperature remains elevated.     

Factors influencing the upper temperature tolerances of three mussel species in a brackish water canal: size, season and laboratory protocols

Mussels are the most problematic organisms encountered in the water intake systems of electrical power plants. Various fouling control measures are adopted, among which heat treatment is considered the relatively more attractive from economic and ecological points of view. Thermal tolerance experiments were carried out to determine the effects of mussel size (2-20 mm shell length), season (breeding vs non-breeding), nutritional status (fed vs non-fed), acclimation temperature (5-25 degrees C) and acclimation salinity (1-35%o) on the mortality pattern of three important mussel species, viz. a freshwater mussel Dreissena polymorpha, a brackish water mussel Mytilopsis leucophaeata and a marine mussel Mytilus edulis under different temperatures (36-41 degrees C). The mussels in the 10 mm size group exposed to 36 degrees C showed 100% mortality after 38 min (D. polymorpha), 84 min (M. edulis) and 213 min (M. leucophaeata). The effect of mussel size on M. edulis and M. leucophaeata mortality at different temperatures was significant, with the largest size group of mussels showing greater resistance, while no significant size-dependence was observed in the case of D. polymorpha. All the three mussel species collected during the non-breeding season (June-October). Nutritional status had no significant influence on the thermal tolerance of the three mussels; fed and non-fed mussels showed 100% mortality at comparable rates. Acclimation temperature had a significant effect on the mortality of all three species. Survival time at any given target temperature increased with increasing acclimation temperature. The acclimation salinity showed no significant effect on the thermal tolerance of the three mussel species. In comparison, M. leucophaeata was more tolerant to high temperature stress than the other two species. The present studies clearly show that various factors can influence the mortality of D. polymorpha, M. edulis and M. leucophaeata to elevated temperatures. The results, therefore, suggest that if heat treatment were to be used as a control measure for these mussels, it has to be employed judiciously, depending on the mussel species, mussel size, breeding season, water temperature and salinity.     

The influence of temperature and dose on antibacterial peptide response against lipopolysaccharide in the blue mussel,Mytilus edulis

Blue mussels (Mytilus edulis) were inoculated with two different doses of lipopolysaccharides (LPS) or phosphate-saline (PS) buffer under different temperature conditions (6 and 20 degrees C). The activity of the antibacterial peptide fraction, purified through reverse phase chromatography from mussel haemolyph, was compared at different time intervals after the inoculation. The activity was determined as the minimal peptide concentration that inhibited growth of the Gram-negative bacteria Escherichia coli D21, by using radial diffusion assay. The antibacterial activity for mussels inoculated with LPS changed over time, both at 6 and 20 degrees C, but those inoculated with PS-buffer did not. The response was enhanced within a time course of 3h. The higher temperature did increase the inhibitory activity and made the mussel respond at an earlier stage, in comparison to that at 6 degrees C. At 20 degrees C, mussels inoculated with 10 microg of LPS responded faster than those inoculated with 0.1 microg of LPS. In addition, cytotoxic effects of LPS on mussel haemocytes were investigated in vitro, using a colorimetric assay. The survival index (SI%) for haemocytes decreased with 76% at 6 degrees C but increased with 100% at 20 degrees C, irrespective of the dose of LPS. This indicated that LPS did not influence the viability of the haemocytes but the high temperature increased their metabolic state. Likely, antibacterial response was provoked by LPS in a dose-dependent manner and favoured by higher metabolic state of the haemocytes, elicited at higher temperature. These results provide important considerations for variability in the internal defence of mussels and consequently, also the retention of viable human pathogens in mussels.     

Dose-response relationship between body temperature and birth defects in radiofrequency-irradiated rats

Five groups of pregnant Sprague-Dawley rats were irradiated for 10-40 min on gestation day 9 in a 27.12-MHz radiofrequency field at a magnetic field strength of 55 A/m and an electric field strength of 300 V/m. The specific absorption rate was 10.8 +/- 0.3 W/kg. Exposures were terminated after the rat's colonic temperature reached 41.0 degrees C, 41.5 degrees C, 42.0 degrees C, 42.5 degrees C, or 43.0 degrees C. A control group was sham irradiated at 0 A/m and 0 V/m on gestation day 9, whereas a second control group was untreated. The incidence of both birth defects and prenatal death was directly related to maternal body temperature once a temperature threshold was exceeded. The temperature threshold for both types of effects was approximately 41.5 degrees C. A few pregnant rats died after exposure to 43.0 degrees C, and higher temperatures were nearly always lethal.     

Effects of cold storage on field and laboratory performance of Trichogramma carverae (Hymenoptera: Trichogrammatidae) and the response of three Trichogramma spp. (T. carverae, T. nr. brassicae, and T. funiculatum) to cold

Delaying emergence of Trichogramma spp. is critical for commercial production. Here, diapause induction was considered for three species (Trichogramma nr. brassicae Bezdenko, Trichogramma carverae Oatman & Pinto, and Trichogramma funiculatum Carver), and the effect of storage temperature (4 degrees C, 8 degrees C, and 10 degrees C) and time (1-8 wk) was investigated for T. carverae. For all species, percentage of emergence was lowered after an initial diapause induction period (28 d at 14 degrees C and a photoperiod of 8:16 [L:D] h) and lowered further after 1-mo storage at 3 degrees C and a photoperiod of 0:24 (L:D) h. No wasps emerged after 2 mo of storage, suggesting that true diapause was not induced. The effect of 1-8-wk storage on wasp quality was investigated for T. carverae both in the laboratory and the field. Initial fieldwork suggested that this species could be successfully stored at 10 degrees C under continuous light (after 5-d development at 25 degrees C and a photoperiod of 16:8 [L:D] h) without reducing the ability of wasps to parasitize eggs in the field. In a second experiment, storage temperatures lower than 10 degrees C and storage times 3 wk or longer had a negative impact on emergence and longevity, and effects were not additive. Negative effects may partly reflect size changes, because size decreased in response to storage time, and there was an interaction between time and temperature effects on size. Storage time was the major factor influencing fecundity and field success; both fitness measures were reduced after storage of 3 wk or longer. T. carverae can therefore be successfully stored for up to 2 wk without detrimental effects, and 10 degrees C is the preferred storage temperature. T. carverae seems to survive unfavorable temperature conditions by entering a state of quiescence.     

In situ infrared spectroscopic investigation of Perna canaliculus mussel larvae primary settlement

The adhesion of Perna canaliculus mussel larvae on a germanium (Ge) prism in filtered seawater at 16 degrees C has been investigated by in situ attenuated total reflection infrared (ATR-IR) spectroscopy. The adhesive from the mussel larvae was spectrally monitored over 2 h and the IR spectrum showed its glycoproteinaceous nature with sulphated and carboxylated moieties. The adhesive from the mussel larvae differs from that of the adults, resembling the mucus secretion found in other benthic marine species at a larval stage. To date, this appears to be the first work describing the main chemical features of secreted adhesive associated with the primary settlement of mussel larvae. The acquired knowledge on the larval adhesive features may lead to enhanced settling methods in aquaculture or to antibiofouling strategies. The ATR-IR approach under temperature control is potentially useful for such studies on other small benthic organisms in both marine and freshwater environments.     

Respiratory response to temperature and hypoxia in the zebra mussel Dreissena polymorpha

The effects of temperature acclimation, acute temperature variation and progressive hypoxia on oxygen consumption rates (VO2) were determined for the zebra mussel Dreissena polymorpha. In the first experiment, after acclimation to 5, 15 or 25 degrees C for at least 2 weeks, VO2 was determined at 5 degrees C increments from 5 to 45 degrees C. VO2 increased in all three acclimation groups from 5 to 30 degrees C, corresponding to the normal ambient temperature range for this species. Mussels displayed imperfect temperature compensation at temperatures above 15 degrees C, but exhibited little acclimatory ability below 15 degrees C. In the hypoxia experiment, VO2 was determined over the course of progressive hypoxia, from full saturation (oxygen tension [PO2]=160 Torr [21.3 kPa]) to a PO2 at which oxygen uptake ceased (<10 Torr [1.3 kPa]). Mussels were acclimated to either 5, 15 or 25 degrees C for at least 2 weeks and their respiratory response to progressive hypoxia was measured at three test temperatures (5, 15 and 25 degrees C). The degree of oxygen regulation increased with increasing test temperature, particularly from 5 to 15 degrees C, but decreased with increasing acclimation temperature. The decreased metabolic rate observed for warm-acclimated animals, particularly in the upper portion of the temperature range of the zebra mussel, may allow for conservation of organic energy stores during warm summer months. Compared to other freshwater bivalves, D. polymorpha is a relatively poor oxygen regulator, corresponding with its preference for well-oxygenated aquatic habitats. In addition, a new quantitative method for determining the degree of oxygen regulation is presented.     

Effect of high-carbon dioxide atmospheres on infestations of apple maggot (Diptera: Tephritidae) in apples

Short-term storage regimens containing elevated atmospheres of carbon dioxide (CO2) were evaluated for their ability to disinfest newly harvested 'McIntosh' apples of apple maggot, Rhagoletis pomonella (Walsh). Infested fruits containing newly laid eggs were either placed directly into the high-CO2 atmosphere at 10 degrees C to expose this life stage, or else held first for 7 d at room temperature, to allow development to the neonate larval stage. Treatment combinations consisted of three different CO2 levels (10.6, 14.9, and 19.0% CO2) and two periods of exposure (7 and 14 d). Apple maggot eggs subjected to the treatments always exhibited some survival, which was lower for the 14-d than the 7-d exposure periods. In contrast, newly hatched larvae were less able to survive the treatments. The 7-d exposure allowed low levels of survival of this life stage, but virtually none survived the 14-d exposure period. To determine the age at which eggs become more susceptible to high-CO2 atmospheres, infested fruits containing eggs three or 3d old were submitted to a 14-d exposure to 19.0% CO2. Survival of 3-d old eggs was similar to that of eggs exposed at an age of 1 d or less, but this dropped to near zero for 5-d old eggs, indicating an increase in susceptibility sometime during the 3-5-d age range. Fruits exposed to 19.0% CO2 for 14 d were significantly firmer than untreated fruits. No apparent browning, internal breakdown or other fruit defects were detected in any of the treatments.     

Variability of the level of thermoresistance as an indicator of the functional status of a cell

Effect of acclimating temperature on the thermoresistance of isolated ciliated gill epithelium of Anodonta anatina was studied. One strip of gills from each mussel was used to determine the initial level of thermoresistance (survival time at 40 degrees C). Other strips were kept at 24 degrees C until their death, and their thermoresistance and fluorescence after Ethidium bromide staining were determined at different time intervals. After a short exposure at 24 degrees C the mean level of thermoresistance of the epithelium increased. The individual shift in the value and direction of cell thermoresistance in different mussels negatively correlated with its initial level. It resulted in narrowing the extent of individual variability of the level of thermoresistance. The prolongation of exposure at 24 degrees C decreased the mean level of thermoresistance, disturbed the negative correlation, and increased the variability of thermoresistance level. The value of cell fluorescence remained constant within 48 h. The increase in fluorescence was observed 12 h after reducing the mean thermoresistance level. Hence, the dynamics of variability in the thermoresistance level is a most sensitive indication of the cell functional state.     

Attachment of the flagellate Giardia lamblia: role of reducing agents, serum, temperature, and ionic composition.

The flagellated protozoan Giardia lamblia has been grown only in highly complex media under reduced oxygen tension. Therefore, the organic and physiological requirements for in vitro attachment and short-term (12-h) survival of this organism were determined. In defined maintenance media, a thiol reducing agent (e.g., cysteine) was absolutely required for attachment and survival of this aerotolerant anaerobe. The crude bovine serum Cohn III fraction greatly stimulated attachment and survival. Attachment was decreased at a reduced temperature (24 degrees C as compared with 35.5 degrees C) and absent at 12 degrees C or below. Attachment and survival were strongly dependent upon pH and ionic strength, with optima at pH 6.85 to 7.0 and 200 to 300 mosmol/kg. Sodium chloride was better tolerated than KC1. Reduction of Ca2+ and Mg2+ to below 10(-8) M did not significantly affect attachment.     

Effects of time and temperature during attachment of sections to microscope slides on immunohistochemical detection of antigens.

To study the effects of time and temperature on attachment of tissue sections to microscope slides, we examined the intensity of immunohistochemical staining of selected antigens in nine different neoplastic and normal tissues after attaching sections at different times and temperatures. Typically, both the temperature and time are minimized when tissue sections attached to slides; however, suboptimal times and temperatures during attachment may result in either loss of tissue due to poor attachment or the necessity for inconvenient staining regimens. Using standard immunohistochemical techniques, 5 microm tissue sections were attached at 58 degrees C for 1, 4 and 24 hr. In a separate study, 5 microm tissue sections were attached for 16 hr at 58, 68 and 80 degrees C. The intensity of staining decreased slightly when the tissue sections were heated at 80 degrees C for 16 hr, but there was little or no decrease when tissues were heated at 68 degrees C or lower for 16 hr, or at 58 degrees C for up to 24 hr.     

Chemical cues from predators and damaged conspecifics affect byssus production in the green-lipped mussel perna viridis

Juveniles of the green-lipped mussel Perna viridis (Bivalvia: Mytilidae) were exposed to either chemical cues of damaged conspecifics or predator, including the swimming crab Thalamita danae and the muricid gastropod Thais clavigera. Byssus production was monitored for 48 h but the highest production rate occurred in the first 6 h. Longer and thicker byssal threads with a larger total volume were produced by P. viridis exposed to damaged conspecifics and predators as compared with the control. Numbers of individuals which shed stalks during the experiment varied significantly from 0 to 50%, and with a significantly higher value obtained for the control. Results indicated that byssus production is a plastic response which could be induced by exposure to chemical signals from predators and damaged conspecifics, the latter producing the largest response. Firmer byssal attachment not only reduces predation risk but also non-predation mortality by securing an animal more effectively to its habitat.     

Thermoregulation during cold exposure: effects of prior exercise.

This study examined whether acute exercise would impair the body's capability to maintain thermal balance during a subsequent cold exposure. Ten men rested for 2 h during a standardized cold-air test (4.6 degrees C) after two treatments: 1) 60 min of cycle exercise (Ex) at 55% peak O(2) uptake and 2) passive heating (Heat). Ex was performed during a 35 degrees C water immersion (WI), and Heat was conducted during a 38.2 degrees C WI. The duration of Heat was individually adjusted (mean = 53 min) so that rectal temperature was similar at the end of WI in both Ex (38.2 degrees C) and Heat (38.1 degrees C). During the cold-air test after Ex, relative to Heat 1) rectal temperature was lower (P < 0.05) from minutes 40-120, 2) mean weighted heat flow was higher (P < 0.05), 3) insulation was lower (P < 0.05), and 4) metabolic heat production was not different. These results suggest that prior physical exercise may predispose a person to greater heat loss and to experience a larger decline in core temperature when subsequently exposed to cold air. The combination of exercise intensity and duration studied in these experiments did not fatigue the shivering response to cold exposure.     

Effects of cooling ovaries before oocyte aspiration on meiotic competence of porcine oocytes and of exposing in vitro matured oocytes to ambient temperature on in vitro fertilization and development of the oocytes

Experiments were conducted to evaluate the effects of cooling porcine ovaries to low temperature (4 degrees C, 15 degrees C, 20 degrees C, 25 degrees C or 30 degrees C) for 1 h on the meiotic competence of their oocytes. Moreover, it was determined whether or not the exposure of in vitro matured oocytes to ambient temperature (20 degrees C, 25 degrees C or 30 degrees C) for 1 h affects the fertilization and developmental competence of the oocytes. There was no difference between the proportions of oocytes that underwent maturation to metaphase II when isolated from control ovaries held at 35 degrees C and ovaries exposed to 30 degrees C. However, the percentages of oocytes from ovaries exposed to 25 degrees C or less were significantly lower than those of oocytes from ovaries exposed to 30 degrees C and control ovaries. The proportions of total and normal fertilization of oocytes that had been exposed to 20 degrees C before in vitro fertilization (IVF) were significantly lower than those of control oocytes maintained at 38.5 degrees C. However, cooling in vitro matured oocytes had no effects on their cleavage and development to blastocysts after IVF. These data suggest that exposing porcine ovaries to a low temperature of 25 degrees C or less before aspiration of oocytes may adversely affect their subsequent in vitro maturation. It may be necessary to maintain the oocytes at a temperature of more than 25 degrees C during manipulation of oocytes for retaining the fertilizability of the oocytes.     

Influence of growth and environmental conditions on cell surface hydrophobicity of Pseudomonas fluorescens in non-specific adhesion

The relative cell surface hydrophobicity (CSH) of 18 soil isolates of Pseudomonas fluorescens, determined by phase exclusion, hydrophobic interaction chromatography (HIC), electrostatic interaction chromatography (ESIC), and contact angle, revealed large degrees of variability. Variation in the adhesion efficiency to Macrophomina phaseolina of the hyphae/sclerotia of these isolates was also examined. Two such isolates with maximum (32.8%; isolate 12-94) and minimum (12%; isolate 30-94) CSH were selected for further study. Early- to mid-log exponential cells of these isolates were more hydrophobic than those in stationary phase, and the CSH of these isolates was also influenced by fluctuations in temperatures and pH. Isolate 12-94 exhibited high CSH (32.3%) at 30 degrees C, compared to lower values (28-24%) in the higher temperature range (35-40 degrees C). Increasing concentrations of either Zn2+, Fe3+, K+, and Mg2+ in the growth medium were associated with the increased CSH. Trypsin, pepsin, and proteinase K (75 to 150 micrograms.mL-1) reduced the CSH of isolate 12-94 cells. CSH was reduced, following exposure to DTT, SDS, Triton X-100, or Tween 80. Prolonged exposure of cells to starvation (60 days) also caused a significant decline in CSH. Several protein bands (18, 21, 23, 26 kDa) of the outer cell membrane were absent in 60-day starved cells compared to unstarved cells. In conclusion, our findings demonstrate that CSH of P. fluorescens isolates may contribute to nonspecific attachment/adhesion onto M. phaseolina hyphae/sclerotia, and the efficiency of adhesion is regulated by growth and other environmental conditions.     

Effect of water temperature on the ability of Diplostomum spathaceum miracidia to establish in lymnaeid snails

The effects of water temperature on the ability of Diplostomum spathaceum miracida to infect and establish patent infections in Lymnaea peregra and L. stagnalis were investigated. Snails were infected over a range of temperatures (6-20 degrees C) and kept thereafter at 20 degrees C or were infected at 20 degrees C and kept at either 14, 20, or 25 degrees C. Infection success was determined after 8 weeks by either observing cercarial shedding or examining snail viscera for sporocysts. The establishment of miracidia declined at lower water temperatures despite maintenance for 8 weeks at 20 degrees C while exposure of snails to miracidia at 20 degrees C and maintenance at different temperatures had little apparent effect. Infection success under these conditions was related more to the numbers of miracidia to which the snails were exposed. However, under this latter experimental regime, the time taken for the infection to become patent clearly depended upon maintenance temperature.