Cloning of heat shock protein genes (hsp70, hsc70 and hsp90) and their expression in response to larval diapause and thermal stress in the wheat blossom midge,Sitodiplosis mosellana

Sitodiplosis mosellana Géhin, one of the most important pests of wheat, undergoes obligatory diapause as a larva to survive unfavorable temperature extremes during hot summers and cold winters. To explore the potential roles of heat shock proteins (hsp) in this process, we cloned full-length cDNAs of hsp70, hsc70 and hsp90 from S. mosellana larvae, and examined their expression in response to diapause and short-term temperature stresses. Three hsps included all signature sequences of corresponding protein family and EEVD motifs. They showed high homology to their counterparts in other species, and the phylogenetic analysis of hsp90 was consistent with the known classification of insects. Expression of hsp70 and hsp90 were highly induced by diapause, particularly pronounced during summer and winter. Interestingly, hsp70 was more strongly expressed in summer than in winter whereas hsp90 displayed the opposite pattern. Abundance of hsc70 mRNA was comparable prior to and during diapauses and was highly up-regulated when insects began to enter the stage of post-diapause quiescence. Heat-stressed over-summering larvae (⩾30 °C) or cold-stressed over-wintering larvae (⩽0 °C) could further elevate expression of these three genes, but temperature extremes i.e. as high as 45 °C or as low as −15 °C failed to trigger such expression patterns. Notably, hsp70 was most sensitive to heat stress and hsp90 was most sensitive to cold stress. These results suggested that hsp70 and hsp90 play key roles in diapause maintenance and thermal stress; the former may be more prominent contributor to heat tolerance and the latter for cold tolerance. In contrast, hsc70 most likely is involved in developmental transition from diapause to post-diapause quiescence, and thus may serve as a molecular marker to predict diapause termination.     

Low-temperature tolerance of false codling moth Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) in South Africa

(1) The low-temperature tolerance of false codling moth (FCM) Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a significant aspect of this pest's population dynamics and has implications for post-harvest control and sterile insect technique programs.

(2) Here, we report results of experiments examining the effects of time, temperature and their interactions on low-temperature tolerance in adult FCM. In addition, using a variety of thermal pre-treatments, we examine the potential for hardening responses over several hours to improve low-temperature tolerance in FCM.

(3) Lower lethal temperature assays showed significant effects of time, temperature and significant interactions between time and temperature on survival (p<0.0001 in all cases). The temperature at which the probability of survival of 50% of the FCM population after 2 h of exposure was −4.5 °C, which varied significantly to −0.5 °C in 10 h. Gender and early adult age did not affect low-temperature tolerance of FCM.

(4) Using a range of non-lethal, low- and high-temperature pre-treatments, FCM survival could not be increased (p>0.84) and thus limited evidence for rapid cold hardening was found. These results are discussed with respect to microclimate temperatures in typical FCM environments and have implications for understanding population dynamics in this species and the diversity of low-temperature responses of insects.

Gene expression of apoptosis-related genes,stress protein and antioxidant enzymes in hemocytes of white shrimp Litopenaeus vannamei under nitrite stress

Apoptotic cell ratio and mRNA expression of caspase-3, cathepsin B (CTSB), heat shock protein 70 (HSP70), manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase (GPx) and thioredoxin (TRx) in hemocytes of white shrimp Litopenaeus vannamei exposed to nitrite-N (20 mg/L) was investigated at different stress time (0, 4, 8, 12, 24, 48 and 72 h). The apoptotic cell ratio and mRNA expression level of CTSB were significantly increased in shrimp exposed to nitrite-N for 48 and 72 h. Caspase-3 mRNA expression level significantly increased by 766.50% and 1811.16% for 24 and 48 h exposure, respectively. HSP70 expression level significantly increased at 8 and 72 h exposure. MnSOD mRNA expression in hemocytes up-regulated at 8 and 48 h, while CAT mRNA expression level increased at 24 and 48 h. GPx expression showed a trend that increased first and then decreased. Significant increases of GPx expression were observed at 8 and 12 h exposure. Expression level of TRx reached its highest level after 48 h exposure. These results suggest that nitrite exposure induces expression of apoptosis-related genes in hemocytes, and subsequently caused hemocyte apoptosis. Meanwhile, expression levels of HSP70 and antioxidant enzymes up-regulated to protect the hemocyte against nitrite stress.     

Differential protein expression for geothermal Agrostis scabra and turf-type Agrostis stolonifera differing in heat tolerance

Heat stress is a major factor limiting the growth of cool-season grasses in warm climatic regions by affecting many physiological processes, including protein metabolism. Protein degradation often occurs with increasing temperatures, but certain specific proteins such as heat shock proteins (HSPs) may be induced or enhanced in their expression under supraoptimal temperatures. The objectives of this study were to determine the critical temperature that causes protein induction or degradation in two Agrostis grass species differing in heat tolerance and to compare protein profiles between the two species under different temperature regimes. Plants of heat-tolerant Agrostis scabra and two cultivars of heat-sensitive Agrostis stolonifera (‘L-93’ and ‘Penncross’) were exposed to constant day/night temperatures of 20, 30, 35, 40, or 45 °C for 14 d. Leaf photochemical efficiency (Fv/Fm), chlorophyll and carotenoid contents, and soluble protein content declined with increasing temperatures. The decreases were the least severe for A. scabra, intermediate for ‘L-93’, and the most severe for ‘Penncross’, indicating interspecific and intraspecific variations in heat tolerance in Agrostis species. Protein degradation was observed at 30–45 °C in both cultivars of A. stolonifera, and at 40–45 °C in A. scabra.HSPs were induced or enhanced at 35–45 °C in ‘L-93’ and A. scabra, and at 40–45 °C in ‘Penncross’. Immunoblotting also revealed stronger expressions of HSP60 and HSP70 in A. scabra or ‘L-93’ than in ‘Penncross’ at 35–45 °C after 3 d. The results suggested the superior heat tolerance of Agrostis grass species and cultivars could be attributed to the early induction of HSPs, particularly small molecular weight (23 kDa), at a lower level of heat stress and the maintenance of protein thermostability, particularly high-molecular weight proteins (83 kDa and large units of Rubisco).     

Effect of thermal stress on HSP70 expression in dermal fibroblast of zebu (Tharparkar) and crossbred (Karan-Fries) cattle

The present studies were conducted to investigate the difference response of dermal fibroblasts to heat stress in Tharparkar and Karan-Fries cattle. Skin is the most important environmental interface providing a protective envelope to animals. In skin, dermal fibroblasts are the most regular cell constituent of dermis that is crucial for temperature homeostasis. The study aimed to examine the reactive oxygen species (ROS) formation, cytotoxicity (%) and heat shock protein 70 (HSP70) genes expression in dermal fibroblast of Tharparkar and Karan-Fries cattle and to assess whether resistance of dermal fibroblast to heat stress is breed specific. Dermal fibroblasts from ear pinna of Tharparkar and Karan-Fries cattle were exposed at 25 °C, 37 °C, 40 °C and 44 °C for 3 h to measure the ROS, cytotoxicity (%) and HSP 70 (HSPA1A, HSPA2 and HSPA8) genes’ expression. The results showed that ROS formation at low temperature (25 °C) decreased in both breeds as compared to control (37 °C) and the differences were significant (P<0.0001). Heat stress at 40 °C did not increase ROS formation significantly in Tharparkar but increased significantly (P<0.001) in Karan-Fries cattle. The overall cytotoxicity (%) was also found to be significantly different (P<0.001) between Tharparkar and Karan-Fries cattle, and on exposure to different temperatures (P<0.001). The cytotoxicity (%) in dermal fibroblast cells of Karan-fries cows was more than Tharparkar. The expression studies indicated that all HSP70 genes (HSPA8, HSPA1A and HSPA2) were up-regulated at different temperatures in both breeds. In Tharparkar, the relative mRNA expression of HSPA8 gene was higher but HSPA1A and HSPA2 genes were low as compared to Karan-Fries cattle. At 40 and 44 °C, the relative expressions of inducible HSP 70 genes (HSPA1A and HSPA2) were higher in Karan-Fries than Tharparkar. In summary, dermal fibroblast resistance to heat shock differed between breeds. Dermal fibroblasts of Tharparkar were observed to be more heat tolerant than crossbred Karan-Fries cattle. The study concludes that zebu cattle (Tharparkar) dermal fibroblasts are more adapted to tropical climatic condition than crossbreed cattle (Karan-Fries). Differences exist in dermal fibroblasts of heat adapted and non-adapted cattle.     

The effect of heat stress on intestinal integrity and Salmonella invasion in broiler birds

The intestinal mucosa works as a barrier to protect the internal environment of the animal from bacteria and bacterial toxins found in the gut lumen. Heat stress may harm this function. Therefore, we designed the current experiment to investigate the effect of heat stress on intestinal integrity, physiological and immunological responses and Salmonella invasion in broiler chickens. At 26 days of age, 72 birds were randomly distributed into 3 treatments, with 8 replicates per treatment and 3 birds per replicate. The three treatments were control treatment; kept at thermoneutral environmental conditions (20 ± 2 °C), chronic heat stress treatment (exposed to 30 ± 2 °C; 24 h/day) and acute heat stress treatment (exposed to 35 ±2 °C from 09:00 to 13:00 and kept at 20 ± 1 °C from 13:00 to 09:00). The heat stress exposure was conducted for 10 successive days. Compared with the control treatment, birds subject to chronic and acute heat stress had reduced (P < 0.05) body weight and body gain and increased (P < 0.05) feed conversion ratio. However, feed intake and mortality rate were only increased (P < 0.05) in the acute heat stress treatment. Rectal temperature and Δ rectal temperature (°C/h) increased (P < 0.05) sharply during the first 2 days of exposure followed by gradual decreases until a plateau was achieved. Heat-stressed birds had increased (P < 0.05) serum concentrations of corticosterone, endotoxin lipopolysaccharide and the systemic inflammatory cytokine: TNF-α and IL-2, as well as a higher (P < 0.05) prevalence of Salmonella spp. in meat and livers, as compared with control treatment. It can be concluded that heat stress impaired intestinal integrity which resulted in increased intestinal permeability to endotoxin, translocation of intestinal pathogens (Salmonella spp.) and serum inflammatory cytokines. Therefore, avoiding thermal dysfunction of intestinal barrier is a significant factor in maintaining welfare, immune status and meat safety of broiler birds.     

Fitness costs associated with different frequencies and magnitudes of temperature change in the butterfly Bicyclus anynana

Plastic responses to changes in environmental conditions are ubiquitous and typically highly effective, but are predicted to incur costs. We here investigate the effects of different frequencies and magnitudes of temperature change in the tropical butterfly Bicyclus anynana, considering developmental (Experiment 1) and adult stage plasticity (Experiment 2). We predicted negative effects of more frequent temperature changes on development, immune function and/or reproduction. Results from Experiment 1 showed that repeated temperature changes during development, if involving large amplitudes, negatively affect larval time, larval growth rate and pupal mass, while adult traits remained unaffected. However, results from treatment groups with smaller temperature amplitudes yielded no clear patterns. In Experiment 2 prolonged but not repeated exposure to 39 °C increased heat tolerance, potentially reflecting costs of repeatedly activating emergency responses. At the same time fecundity was more strongly reduced in the group with prolonged heat stress, suggesting a trade-off between heat tolerance and reproduction. Clear effects were restricted to conditions involving large temperature amplitudes or high temperatures.     

Thermal acclimation in clownfish: An integrated biomarker response and multi-tissue experimental approach

The effects of increased temperature were tested in Amphiprion ocellaris, using a cellular diagnostics approach (in several tissues) combined with an organismal approach (body condition). Clownfish were exposed to a one month experiment following two temperature treatments: control (26 °C) and elevated temperature (30 °C). Fish were sampled at 0, 7, 14, 21 and 28 days for (1) assessment of stress biomarkers (catalase, lipid peroxidation, glutathione-S-transferase, superoxide dismutase, acetylcholinesterase, heat shock protein 70 kDa and ubiquitin – in brain, gills, liver, intestine and muscle), (2) estimation of integrated biomarker response index based on the biomarkers tested and (3) assessment of Fulton’s K index. Results show all biomarkers except acetylcholinesterase responded consistently and significantly to elevated temperature across tissue types suggesting they are suitable indicators of thermal stress in A. ocellaris. Biomarker levels were tissue-specific, and in addition, the most reactive tissues to temperature were muscle, gills and liver which suggest that highly oxygenated tissues seem to be the most responsive under thermal stress. The most responsive sampling times to increased temperature were T7 and T28: thermal stress was observed after 7 days of exposure (biomarker levels increased), then a pattern of decrease in biomarker levels towards the end of the experiment was observed, which may suggest fish were able to acclimate to exposure conditions. This indicates that A. ocellaris probably lives far from its upper thermal limit and is capable of adjusting the protein quality control system and enzymes’ activities to protect cell functions under elevated temperatures. The temperature treatment did not significantly influence body condition of the animals but biomarkers were negatively correlated to wet body weight. This suggests that thermal acclimation incurs at some energetic cost. In conclusion, these results suggest that this coral reef fish species presents a significant acclimation potential under ocean warming scenarios of +4 °C.     

Whole-plant carbon relations and root respiration associated with root tolerance to high soil temperature for Agrostis grasses

Plant tolerance to high soil temperature may be related to the adjustment in carbon production and utilization. The objective of this study was to determine changes in whole-plant carbon balance and root respiration rate in relation to root tolerance to high soil temperature for two Agrostis grass species varying in heat tolerance. Plant tolerance to high soil temperature was compared between Agrostis scabra, a thermal grass species adapted to chronic high-temperature soils in the geothermal areas in Yellowstone National Park, and creeping bentgrass (Agrostis stolonifera), a cultivated grass species adapted to cool climatic regions. Plant roots were exposed to low soil temperature (20 °C) or high soil temperature (37 °C) for 17 days in water baths placed in a controlled-environment growth chamber. Root biomass and cell membrane stability were determined to evaluate root thermotolerance of both species. Canopy photosynthetic rate (Pn), whole-plant respiration rate, root respiration rate, and total non-structural carbohydrate (TNC) content were measured to assess changes in carbon production and utilization in response to high soil temperature. Root biomass and cell membrane stability declined with increasing soil temperature, but the decline was much less for A. scabra than A. stolonifera, suggesting that roots of A. scabra were more tolerant to heat stress. Canopy Pn decreased and whole-plant respiration rate increased for A. stolonifera, but canopy Pn and respiration rate were unchanged for A. scabra in response to increasing soil temperature. After 17 days of high soil temperature treatment, A. stolonifera exhibited carbon deficit at the whole-plant level, whereas A. scabra maintained positive carbon gain. Root respiration of plants previously grown at 20 °C increased after a short-term treatment (24 h) at 37 °C, but the increase was significantly lower for A. scabra than for A. stolonifera. TNC content in roots did not show response to short-term (24 h) changes in temperature and did not exhibit species variations. Leaves of A. scabra, however, maintained TNC content under both low and high temperature regimes. Our results suggest that root thermotolerance of cool-season grasses could be related to the maintenance of positive whole-plant carbon balance, and down-regulation of whole-plant and root respiration rates in response to increasing soil temperature.     

Estrus related differences in response to a hot environment in telemetry-equipped female rats

The purpose of this study was to quantitate the physiological responses of female rats during estrus (E) and non-estrus (NE) phases of their cycle to a short-term exposure to a hot environment (38°C, 90 min). Heart rate (HR), core temperature (T_c), activity level (ACT), and evaporative water loss (EWL) responses in telemetry-implanted rats (n=9) in the heat were compared to responses at room temperature (23°C, 90 min). Both at room temperature and in the heat, T_c changed significantly across time and was significantly higher in the heat. At room temperature, HR was not different between estrus stages or across time while in the heat HR changed significantly across time. ACT declined for 20 min and then remained similar among groups for the duration of the exposure. EWL was greater in the heat than at room temperature although during both exposures EWL did not change significantly across time or between stages. These results indicate that in rats the reproductive stage does not affect their response to short-term environmental stimuli.     

Mitigating thermal effect of behaviour and microhabitat on the intertidal snail Littorina saxatilis (Olivi) over summer

High shore intertidal ectotherms must withstand temperatures which are already close, at or beyond their upper physiological thermal tolerance. Their behaviour can provide a relief under heat stress, and increase their survival through thermoregulation. Here, we used infrared imaging to reveal the thermoregulatory behavioural strategies used by the snail Littorina saxatilis (Olivi) on different microhabitats of a high shore boulder field in Finistère (western France) in summer. On our study site, substrate temperature is frequently greater than L. saxatilis upper physiological thermal limits, especially on sun exposed microhabitats. To maintain body temperatures within their thermal tolerance window, withdrawn snails adopted a flat posture, or elevated their shells and kept appended to the rock on the outer lip of their aperture with dried mucous (standing posture). These thermal regulatory behaviours lowered snail body temperatures on average by 1–2 °C. Aggregation behaviour had no thermoregulatory effect on L. saxatilis in the present study. The occupation of biogenic microhabitats (barnacles) was associated with a 1 °C decrease in body temperatures. Barnacles and microhabitats that experienced low sun exposure, low thermal fluctuations and low thermal maxima, could buffer the heat extremes encountered at high shore level especially on sun exposed microhabitats.     

A proteomic analysis of the aphid Macrosiphum euphorbiae under heat and radiation stress

Temperature and solar radiation can be important sources of abiotic stress for small herbivorous insects living in close association with plants. We examined the effects of daily fluctuations of heat and UV radiation on the proteome and performance of winged and wingless morphs of the aphid Macrosiphum euphorbiae. A daily regime of 4 h of heat stress at 35 °C had more negative effects on the aphid's fitness than a similar period of UV-B stress (11.6 kJ m^?2 per day), and these effects were most pronounced on wingless aphids. Aphid proteomes as detected on 2-D gels revealed ～470 protein spots, with the fluctuating heat stress leading to many more changes than exposure to UV-B. The reduced performance of aphids under heat stress correlated with lower abundance of several enzymes in central pathways of energy metabolism, including the TCA cycle and the respiratory chain. Several exoskeletal proteins were induced or their abundance was increased under high temperature stress, suggesting that cuticle barrier enhancement at molting in response to heat stress is an aphid adaptation to stressful thermal conditions. The proteome of winged aphids was more broadly modulated under stress than that of wingless aphids. Greater homeostatic capabilities as revealed at the proteomic level could explain the higher tolerance of the alate aphid morph to environmental stress and its more stable performance and fitness.     

Temperature-dependent physiological and biochemical responses of the marine medaka Oryzias melastigma with consideration of both low and high thermal extremes

This study aimed to investigate temperature effect on physiological and biochemical responses of the marine medaka Oryzias melastigma larvae. The fish were subjected to a stepwise temperature change at a rate of 1 °C/h increasing or decreasing from 25 °C (the control) to six target temperatures (12, 13, 15, 20, 28 and 32 °C) respectively, followed by a 7-day thermal acclimation at each target temperature. The fish were fed ad libitum during the experiment. The results showed that cumulative mortalities were significantly increased at low temperatures (12 and 13 °C) and at the highest temperature (32 °C). For the survivors, their growth profile closely followed the left-skewed ‘thermal performance curve’. Routine oxygen consumption rates of fish larvae were significantly elevated at 32 °C but suppressed at 13 and 15 °C (due to a high mortality, larvae from 12 °C were not examined). Levels of heat shock proteins and activities of malate dehydrogenase and lactate dehydrogenase were also measured in fish larvae exposed at 15, 25 and 32 °C. The activities of both enzymes were significantly increased at both 15 and 32 °C, where the fish larvae probably suffered from thermal discomfort and increased anaerobic components so as to compensate the mismatch of energy demand and supply at these thermal extremes. Coincidently, heat shock proteins were also up-regulated at both 15 and 32 °C, enabling cellular protection. Moreover, the critical thermal maxima and minima of fish larvae increased significantly with increasing acclimation temperature, implying that the fish could develop some degrees of thermal tolerance through temperature acclimation.     

Stage- and sex-specific heat tolerance in the yellow dung fly Scathophaga stercoraria

Thermal tolerance varies at all hierarchical levels of biological organization: among species, populations, individuals, and even within individuals. Age- or developmental stage- and sex-specific thermal effects have received relatively little attention in the literature, despite being crucial for understanding thermal adaptation in nature and responses to global warming. We document stage- and sex- specific heat tolerance in the yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae), a species common throughout the northern hemisphere that generally favours cool climates. Exposure of eggs to temperatures up to 32 °C did not affect larval hatching rate, but subsequent egg-to-adult survival at a benign temperature was reduced. Permanent transfer from benign (18 °C) to hot temperatures (up to 31 °C) at different larval and pupal stages strongly decreased egg-to-adult survival, though survival continuously improved the later the transfer occurred. Temporary transfer for only two days increased mortality more weakly, survival being lowest when temperature stress was imposed early during the larval or pupal stages. Adult flies provided with sugar and water tolerated 31 °C longer than previously thought (5 days in males to 9 days in females). Eggs were thus less susceptible to thermal stress than larvae, pupae or adults, in agreement with the hypothesis that more mobile stages require less physiological protection against heat because they can behaviourally thermoregulate. The probability of mating, of laying a clutch, and hatching success were generally independently reduced by exposure of females or males to warm temperatures (24 °C) during the juvenile or adult stages, with some interactions evident. High temperature stress thus affects survival differentially depending on when it occurs during the juvenile or the pre-reproductive adult life stage, and affects reproductive success via the mating behaviour of both sexes, female physiology in terms of oviposition, and fertility via sperm and/or egg quality. Our results illustrate that temperature stress, even when moderate and temporary, during early development can have profound lethal and non-lethal fitness-consequences later in life.     

Effect of in vitro zinc supplementation on HSPs expression and Interleukin 10 production in heat treated peripheral blood mononuclear cells of transition Sahiwal and Karan Fries cows

The changing climatic scenario with apprehended rise in global temperature is likely to affect the livestock adversely vis-à-vis production and reproduction. This has prompted more focus in addressing the unfavorable effects of thermal stress in livestock system. Presuming that the trace element zinc is indispensible for cellular antioxidant system and immune function, the present study was designed to investigate the effect of zinc treatment on heat stress alleviation and immune modulation in peripheral blood mononuclear cells (PBMC) of indigenous and crossbred transition cows. Twelve cows, six each of Sahiwal and Karan Fries (KF) in their second parity with confirmed pregnancy were selected for the experiment. The blood samples were collected at −21, 0 and +21 days in relation to expected date of calving. The experiment was carried out in vitro after isolating PBMC from whole blood. The 48 h cultured PBMC were subjected to assorted levels of exposures viz. 37 °C, 42 °C to impose heat stress and 42 °C+zinc to alleviate heat stress and modulate immunity. The PBMC viability was 86%, 69% and 78%, respectively. The mRNA expression of heat shock proteins (HSP 40, 70 and 90α) and Interleukin-10 (IL-10) production varied between the two breeds vis-à-vis days and levels of exposure. The mRNA expression of HSP40 and HSP70 was significantly (P<0.05) higher in Karan Fries than the Sahiwal cows. Both the breeds showed maximum expression of HSP on the day of parturition, more so in KF than Sahiwal. There was a significant (P<0.05) difference in the HSP mRNA expression at different levels of exposure. Zinc treatment to heat stressed PBMC caused a significant (P<0.05) down regulation of HSP. For immune status, anti-inflammatory cytokine, IL-10 in the culture supernatant was accessed. The IL-10 was significantly (P<0.05) higher in Karan Fries (168.18±14.09 pg/ml) than the Sahiwal cows (147.24±11.82 pg/ml). The IL-10 concentration was highest on the day of calving. Zinc treatment reduced the IL-10 concentration. From the study, it could be concluded that the zinc supplementation in heat stressed PBMC can ameliorate thermal stress and modulate immune response which can act as a model for reducing heat stress during the periparturient period in tropical livestock.     

Exposure of mussels to a polluted environment: Insights into the stress syndrome development

Coastal environments are often subjected to contamination, whose biological impact is profitably evaluated through sentinel organisms and biomarkers. mRNA profiling was also proposed as a potential biomarker, whose relevance is still under discussion. Indeed, correlation between molecular and cell-organism responses need further investigations, especially under field conditions. In this study, we followed the development of physiological alterations in Mytilus galloprovincialis transplanted into a polluted coastal lagoon for 2, 4, 7, 14 and 30 days. Three consolidated biomarkers were measured, i.e. lysosomal membrane stability, lipofuscin and metallothionein contents. In parallel, the expressions of stress-related genes encoding metallothioneins (mt10 and mt20), 70-kDa heat shock proteins (MgHSC70 and MgHSP70), and Multi Xenobiotic Resistance-related transporters (MgPgp, MgMrp2, and MgMvp) were analyzed, to have a greater insight into the time-related evolution of the response. Significant (p < 0.05) biomarker responses were induced after 7 days of exposure and further increased with time, whereas gene expression profiles were dramatically altered 2 days after transplanting. Biomarkers and gene expression profiles indicated a stress syndrome development in mussels, although with different temporal patterns. Their combined application provided insights into the molecular and cellular basis of mussel responses to challenging environments, and may have far-reaching implications for monitoring environmental health.