Heat shock protein induction in the freshwater prawn Macrobrachium malcolmsonii: acclimation-influenced variations in the induction temperatures for Hsp70

The intracellular build-up of thermally damaged proteins following exposure to heat stress results in the synthesis of heat shock proteins (Hsps). In the present study, the upper thermal tolerance and expression of heat shock protein 70 (Hsp70) were examined in juveniles of the freshwater prawn Macrobrachium malcolmsonii that had been acclimated at two different temperatures, i.e. 20 degrees C (group A) and 30 degrees C (group B), in the laboratory for 30 days. Upper thermal tolerance was determined by a standard method. For heat-shock experiments, prawns in groups A and B were exposed to various elevated temperatures for 3 h each, followed by 1 h recovery at the acclimation temperature. Endogenous levels of Hsp70 were determined in the gill, heart, hepatopancreas and skeletal muscle tissues by Western blotting analysis of one dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The critical thermal maximum (CT max) for prawns in groups A and B was 37.7+/-0.27 degrees C and 41.41+/-0.16 degrees C, respectively. In general, Western blotting analysis for Hsp70 revealed one band at the 70 kDa region, containing both constitutive (Hsc70) and inducible (Hsp70) isoforms, in the gill and heart tissues; these were not detected in the hepatopancreas and skeletal muscle tissues. The onset temperature for Hsp70 induction in both gill and heart tissues was 30 degrees C for prawns in group A and 34 degrees C for those in group B. The optimum induction temperatures (at which Hsp70 induction was maximum) were found to be 34 degrees C and 32 degrees C, respectively, in the gill and heart tissues of group A prawns, and 38 degrees C and 36 degrees C, respectively, for group B prawns. These results suggest that the temperature at which acclimation occurs influences both upper thermal tolerance and Hsp70 induction in M. malcolmsonii.     

Effect of exposure to cadmium on the tropical freshwater prawn Macrobrachium rosenbergii

The effect of cadmium (Cd) on mortality, resistance and bioaccumulation in the tropical freshwater prawn Macrobrachium rosenbergii from Egypt were studied. Survival of prawns exposed to Cd doses over 60 μg l^?1 were significantly lower than that of prawns exposed to lower doses. After 96 h prawns exposed to >40 μg l^?1 of Cd had a greater reduction in total haemocyte count and phagocytic activity than those exposed to lower concentrations. Bioaccumulation of Cd in the gills, hepatopancreas and muscles was variable. Cadmium accumulated significantly in the gills and hepatopancreas, but Cd accumulation in the muscles increased only marginally. Macrobrachium rosenbergii manifested histopathological alterations in the gills, hepatopancreas and muscles when exposed to various concentrations of cadmium.     

Lead toxicity at different life stages of the giant prawn (Macrobrachium rosenbergii,de Man): considerations of osmoregulatory capacity and histological changes in adult gills

This study evaluated the acute toxicity of lead in different life stages of the freshwater prawn Macrobrachium rosenbergii, determined the effect of sublethal Pb concentrations on osmoregulatory capacity (OC), measured the Pb level in gills, and investigated the effect of Pb on the structure of the gills of adult prawns. The 24-, 48-, and 96-h LC₅₀ values for Pb to M. rosenbergii increased progressively with increasing life stage, from post-larvae (PL), juvenile to adult. The 24- and 48-h LC₅₀ values for post-larvae ranged from 0.01- to 0.09-mg Pb/L. The 24-, 48-, and 96-h LC₅₀ values for Pb were lower at 12 ppt than those at 0 ppt for either the juveniles or the adults. At 12 ppt, the 96-h LC₅₀ values in PL11, juvenile and adult were 0.47-, 0.58-, and 2.03-mg Pb/L, respectively. Meanwhile, at 12 ppt, the 96-h LC₅₀ values in PL11, juvenile and adult were 0.63-, 4.44-, and 7.98-mg Pb/L, respectively. In adults, the OC values of controls and prawns exposed to 2- and 4-mg Pb/L at 0 ppt were not significantly different. The OC of prawns exposed to and 2-mg Pb/L at 12 ppt increased by 72 and 109% from the OC of the control prawns. At media 12 ppt, the OC value of prawns exposed to 1-mg Pb/L was significantly different from that of prawns exposed to 2-mg Pb/L. The concentrations of Pb in gill tissues increased significantly in Pb exposed prawns both at 0 and 12 ppt. The level of Pb in gills of prawns exposed to 2-mg Pb/L at 12 ppt was not significantly different from those exposed at 0 ppt. The severe toxic actions of Pb were noted in gills of prawns exposed in media 12 ppt. Hyperplasia and necrosis were observed in gill lamellae, resulting in abnormal gill tips after Pb exposure at media 12 ppt. Since the effect of Pb is more pronounced in higher salinity (12 ppt) than in freshwater (0 ppt) it is clear that aquaculture of M. rosenbergii should be conducted in freshwater ponds.     

Cytochrome P450-dependent monooxygenase system mediated hydrocarbon metabolism and antioxidant enzyme responses in prawn, Macrobrachium malcolmsonii

We investigated the alteration of cytochrome P450-dependent monooxygenase enzymes and antioxidant enzymes in response to oil effluent in freshwater prawn, Macrobrachium malcolmsonii. The prawns were exposed to two sublethal (10% [0.91 ppt] and 25% [2.3 ppt] of 5-day median lethal concentration) concentrations of oil. After 30 days, treated prawns were transferred into untreated freshwater and depuration was followed for another 30 days. At 7-day intervals, hydrocarbons and detoxifying enzymes were analysed in the hepatopancreas. Accumulation of hydrocarbon in the tissues gradually increased when exposed to sublethal concentration of oil effluent associated with enhanced levels of cytochrome P450, NADPH cytochrome c reductase and cytochrome b(5). During depuration, the levels of accumulated hydrocarbons decreased due to the induction of these detoxifying enzymes. Oil derived hydrocarbon mediated oxyradical production would have occurred in M. malcolmsonii. This was confirmed by elevated levels of superoxide dismutase (SOD) and catalase (CAT). Thus, cytochrome P450-dependent monooxygenase enzymes and antioxidant enzymes in oil-exposed prawns demonstrate a well-established detoxifying mechanism in M. malcolmsonii.     

Effect of cadmium on survival,osmoregulation and gill structure of the Sunda prawn,Macrobrachium sintangense (de Man), at different salinities

The prawn Macrobrachium sintangense is likely to be subjected to occasional exposure to combined metal and saline stressors in its natural environment. This research evaluated the acute toxicity (96?h LC₅₀) of cadmium (Cd) on the prawn M. sintangense, with respect to the osmoregulatory capacity (OC) of prawns and to document histological changes in the gills after exposure to sublethal Cd concentrations at different salinities. The 96?h LC₅₀ of Cd to M. sintangense decreased with increasing salinity. The 96?h LC₅₀ values were 89.12 (72.53–109.50), 681.26 (554.20–837.46) and 825.37 (676.99–1006.27) μg CdL^?1 at 0, 10 and 20 ppt, respectively. The OC of prawns exposed to 30?μg?CdL^?1 at 0 ppt and to 300?μg?CdL^?1 at10 ppt decreased significantly compared with that of control prawns exposed to 0 and 10 ppt respectively. Swelling, hyperplasia and necrosis of gill lamellae resulting in the loss of marginal canals were observed in the gills of prawns exposed to 30?μg?CdL^?1 at 0 ppt and to 300?μg?CdL^?1 at 10 ppt for 7?days.     

Induction of metallothionein-like proteins by mercury and distribution of mercury and selenium in the cells of hepatopancreas and gill tissues in mussel Mytilus galloprovincialis

The binding of mercury (Hg) to metallothioneins (MTs) and the relation between Hg and selenium in supernatants of hepatopancreas and gill tissues of the common mussel Mytilus galloprovincialis (Lamarck, 1819) was investigated. The mussels were exposed to different Hg concentrations in laboratory conditions: 2.5 μgHg/L, 4 d exposure (short term) and 60 μgHg/L, 33 d exposure (long term). In addition, the results were compared to those found for mussels from nature (polluted and unpolluted region). In control and short-term-exposed mussles, the level of Hg extraction (cytosol) from hepatopancreas and gill cells was very low with respect to the total Hg concentrations in the corresponding tissues, around 10% in control and around 20% after exposure. As expected, Hg exposure was followed by Se increase. For Se, the levels of extraction were higher, around 20% in control and up to 50% (heaptopacrease) of 70% (gills) after exposure. In order to study the distribution of Hg and Se in the cells of these organs, the total Hg and Secconcentrations were analyzed in the subcellular fractions obtained after differential centrifugation. Although after exposure the concentrations of both element increased in all subcellular fractions, their percentages in particular fractions were lower or higher. In this study, the convincing binding of Hg to metallothionein-like proteins was perceived after long-term laboratory exposure (gills, heapatopancreas) and in wild mussels collected near industrial port (hepatopancreas). In latter case, we also detected the traces of Se bound to the MT fractions after size-exclusion chromatography.     

Methylparathion, carbaryl and aldrin impact on nitrogen metabolism of prawn, Penaeus indicus

Changes in hepatopancreas, muscle and gill tissue nitrogen metabolic profiles were studied in a penaeid prawn, Penaeus indicus, following its exposure to sublethal concentrations of methylparathion, carbaryl and aldrin. In all the insecticide exposed prawn tissues, Ammonia levels were significantly increased and a shift in the nitrogen metabolism towards the synthesis of urea and glutamine was observed. Inhibition of glutamate oxidation to ammonia and alpha-ketoglutarate by glutamate dehydrogenase suggests a mechanism whereby hyperammonemia is reduced by minimizing the addition of further ammonia to the already existing elevated ammonia pool. Increased alanine and aspartate aminotransferases demonstrates the onset of gluconeogenesis. Mechanisms to detoxify the ammonia by enhancing the synthesis of urea and glutamine at the cellular level was observed in the selected tissues pave way for the survivability of prawns in insecticide polluted environs.     

重金属离子对凡纳滨对虾肝胰脏、鳃丝和血液SOD活力的影响

研究了3种重金属离子(Cu²⁺、Zn²⁺、Cd²⁺)在96 h内对凡纳滨对虾(Litopenaeus vannamei)对肝胰脏、鳃丝和血液超氧化物歧化酶(SOD)活力的影响.结果表明,凡纳滨对虾SOD活力在3种重金属离子作用下随取样时间变化显著(P＜0.0),Cu²⁺在实验浓度范围内(0.1～1 mg·L^-1),肝胰脏、鳃丝和血液的SOD活力随时间延长呈一峰值变化,Zn²⁺在10 mg·L^-1时对肝胰脏表现为显著抑制作用,Cd²⁺在0. mg·L^-1时对肝胰脏和鳃丝起显著抑制作用,0.2 mg·L^-1对鳃丝SOD活力无显著变化(P＞0.0),其他浓度Zn²⁺(＜10 mg·L^-1)、Cd²⁺(＜0.2 mg·L^-1)对各组织器官SOD活力的影响随时间延长均呈现先升高后下降的趋势.3种重金属离子对凡纳滨对虾肝胰脏、鳃丝、血液SOD活力的影响呈现明显的剂量-时间效应关系.其SOD活力大小顺序为肝胰脏＞鳃丝＞血液,3种重金属离子对凡纳滨对虾伤害大小顺序为Cd²⁺＞Cu²⁺＞Zn²⁺.     

Lack of glucose and hsp70 responses in haddock Melanogrammus aeglefinus (L.) subjected to handling and heat shock

Juvenile haddock Melanogrammus aeglefinus ( c. 39 g) were exposed to either a handling stressor (1 min out of water) or heat shock (increase from 10 to 15° C for 1 h), and plasma cortisol, plasma glucose and gill hsp70 levels were determined before, and at 1, 3, 6, 12, 24 and 48 h post-stress. The pattern of cortisol increase was similar following both stressors, with levels increasing by 25-fold at 1 h post-stress, but returning to pre-stress levels (2–5 ng ml⁻¹) by 3 h. In contrast, neither handling nor heat shock caused an increase in plasma glucose levels. Although gill hsp70 was detected, presumably constitutive levels, in both control and heat shocked groups, there were not significant changes in gill hsp70 levels after exposure to heat shock. The lack of glucose and hsp70 responses to these typical stressors is consistent with previous studies on Atlantic cod Gadus morhua , and suggests that the stress physiology of Gadidae differs from the 'typical' teleost.     

Hyperglycaemic effects of 5-hydroxytryptamine and dopamine in the freshwater prawn, Macrobrachium malcolmsonii

The effects of 5-hydroxytryptamine (5-HT; serotonin) and dopamine (DA) on tissue carbohydrate metabolism and haemolymph glucose levels in the freshwater prawn, Macrobrachium malcolmsonii, were investigated. Injection of 5-HT and DA produced hyperglycaemia in a dose-dependent and time-dependent manner, with DA being more effective than 5-HT. Interestingly, 5-HT and DA induced hyperglycaemia only in intact prawns but not in bilaterally eyestalk-ablated individuals. Total carbohydrate (TCHO) and glycogen levels decreased and phosphorylase activity increased in the hepatopancreas and muscle of intact prawns after being injected with 5-HT or DA. However, bilateral eyestalk ablation decreased haemolymph glucose and tissue phosphorylase activity and increased TCHO and glycogen levels of the hepatopancreas and muscle. Injection of 5-HT or DA did not cause significant changes in these variables in eyestalk-ablated prawns. It is hypothesized that 5-HT and DA induce hyperglycaemia in prawns by stimulating the release of crustacean hyperglycaemic hormone (CHH) from the X-organ sinus gland (XO-SG) complex located in the eyestalk.     

Dietary vitamin-E modulates antioxidant defence system in giant freshwater prawn, Macrobrachium rosenbergii

The objectives of the present study were to determine the effect of supplementary vitamin-E (200, 400 and 600 mg/kg feed) on lipid peroxidation (LPX) and antioxidant defence system in gills and hepatopancreas of the freshwater prawn, Macrobrachium rosenbergii. Results indicated that vitamin-E inhibited LPX in the hepatopancreas in a comparatively lower dose than gills. Superoxide dismutase (SOD) activity was decreased significantly in gills in response to all the three supplemented diet, but in hepatopancreas decrease was observed only in response to higher doses of vitamin-E (400 and 600 mg/kg feed). Catalase (CAT) activity was reduced significantly only in gills but not in hepatopancreas. While glutathione peroxidase (GPX) activity was significantly elevated in the hepatopancreas by vitamin-E, its activity remains unaltered in gills. On the contrary, glutathione reductase (GR) activity was decreased in gills but that of hepatopancreas was constant. Glutathione (GSH) content of both gills and hepatopancreas was substantially elevated in the vitamin-E supplemented prawns. Although the ascorbic acid (ASA) content of gills was unchanged by vitamin-E, its level elevated significantly in hepatopancreas. Thus the findings of the present investigation suggest that dietary vitamin-E is capable of reducing LPX level and can modulate antioxidant defence system in gills and hepatopancreas, nevertheless, the response is highly tissue specific. It is further observed that highest dose of vitamin-E (600 mg/kg feed) could not render much additional protection in both the tissues.     

Waterborne vs. dietary copper uptake in rainbow trout and the effects of previous waterborne copper exposure

Juvenile rainbow trout (Oncorhynchus mykiss) were exposed to waterborne Cu (22 microg/l) in moderately hard water for up to 28 days. Relative to control fish kept at background Cu levels (2 microg/l), Cu-preexposed fish displayed decreased uptake rates of waterborne Cu via the gills but not of dietary Cu via the gut during 48-h exposures to (64)Cu-radiolabeled water and diet, respectively. At normal dietary and waterborne Cu levels, the uptake rates of dietary Cu into the whole body without the gut were 0.40-0.90 ng. g(-1). h(-1), >10-fold higher than uptake rates of waterborne Cu into the whole body without the gills, which were 0.02-0.07 ng. g(-1). h(-1). Previously Cu-exposed fish showed decreased new Cu accumulation in the gills, liver, and carcass during waterborne (64)Cu exposures and in the liver during dietary (64)Cu exposures. A 3-h gill Cu-binding assay showed downregulation of the putative high-affinity, low-capacity Cu transporters and upregulation of the low-affinity, high-capacity Cu transporters at the gills in Cu-preexposed fish. Exchangeable Cu pools in all the tissues were higher during dietary than during waterborne (64)Cu exposures, and previous Cu exposure reduced waterborne exchangeable Cu pools in gill, liver, and carcass. Overall, these results suggest a quantitatively greater role for the dietary than for the waterborne route of Cu uptake, a key role for the gill in Cu homeostasis, and important roles for the liver and gut in the normal metabolism of Cu in fish.     

The effect of time on physiological changes in eel Anguilla anguilla, induced by lindane.

1. Eel were exposed to a sublethal concentration of lindane (0.335 ppm) for 6, 12, 24, 48, 72 and 96 hr. 2. Concentrations of glycogen, glucose, lactate, pyruvate and lipids were determined in gill tissue after lindane exposure. 3. Gill glycogen decreased and glucose levels increased at 6 hr of treatment, lactate and pyruvate concentration increased between 6 and 48 hr. Total lipid values decreased between 6 and 24 hr; thereafter, the levels increased up to 72 hr of exposure. 4. Clear changes were found in all parameters tested in gill tissues. The observed effects of lindane on metabolism in fish are discussed in relation to acute stress syndrome.     

中华绒螯蟹对Pb和Cd的富集与释放特性

应用生物富集双箱动力学模型模拟了中华绒螯蟹(Eriocheir sinensis)分别在Pb浓度为0.25、0.50、0.75mg/L,Cd浓度为0.025、0.050、0.075mg/L的单一水环境中暴露时,蟹鳃、肝胰腺、肌肉和血淋巴对Pb和Cd的生物富集与释放实验,并通过非线性拟合得到中华绒螯蟹对Pb和Cd的富集速率常数k1、排出速率常数k2、生物富集系数BCF、生物半衰期B1/2、富集平衡时生物体内Pb和Cd含量CAmax等动力学参数。结果表明:(1)中华绒螯蟹对Pb和Cd具有明显的富集,蟹鳃、肝胰腺和肌肉中Pb和Cd的含量与富集时间和水环境中Pb和Cd暴露浓度表现出了很好的正相关,血淋巴在富集阶段没有明显的规律。理论平衡状态下鳃、肝胰腺和肌肉中Pb和Cd含量CAmax随着暴露浓度的增大而增大,且成正相关。(2)Pb和Cd在中华绒螯蟹组织器官中的富集具有选择性,开始实验前,Pb在中华绒螯蟹体内的的分布规律为:肝胰腺>鳃>肌肉>血淋巴;Cd的分布规律为:鳃>肝胰腺>血淋巴>肌肉。在实验浓度的Pb和Cd水环境中暴露16d后,Pb的分布规律为:鳃>肝胰腺>肌肉>血淋巴;Cd的分布规律为:肝胰腺>鳃>肌肉>血淋巴。(3)中华绒螯蟹对Pb和Cd的生物富集和释放都较缓慢。经过16d的生物富集,各组织器官中Pb和Cd的含量均未达到稳态平衡。Pb和Cd在组织器官中的生物富集系数(BCF)范围分别为5-51和6-3148,中华绒螯蟹对Cd的富集能力明显高于Pb(P1/2)范围分别为4-9d和8-57d,中华绒螯蟹对Cd的排出能力明显低于Pb。       

Impairment of the stress response in matrinxã juveniles (Brycon amazonicus) exposed to low concentrations of phenol

In this study we measured plasma cortisol, plasma glucose, plasma sodium and potassium, and liver and gill hsp70 levels in juvenile matrinxã (Brycon amazonicus) subjected to a 96 h exposure to phenol (0, 0.2, and 2.0 ppm), and the effect of this exposure on their ability to respond to a subsequent handling stress. Fish were sampled prior to initiation of exposure and 96 h, and at 1, 6, 12, and 24 h post-handling stress. During the 96 h exposure, plasma cortisol and glucose levels remained unchanged in all treatments. While plasma sodium levels were significantly reduced in all groups, plasma potassium levels only decreased in fish exposed to 0 and 0.2 ppm of phenol. Liver hsp70 levels decreased significantly at 96 h in fish exposed to 2.0 ppm of phenol. All groups, except fish exposed to 0.2 ppm of phenol, were able to increase plasma cortisol and glucose levels after handling stress. Fish exposed to 2.0 ppm of phenol showed decreased gill and liver hsp70 levels after the handling stress. Our data suggest that exposure to phenol may compromise the ability of matrinxã to elicit physiological responses to a subsequent stressor.     

基于PCR-DGGE基因指纹的对虾体内优势细菌组成分析

采用不依赖分离培养的16S rDNA的PCR-DGGE基因指纹技术对刀额新对虾与中国对虾的鳃部与肠道优势细菌种群组成进行比较分析。研究发现:对虾鳃部与肠道存在着丰富多样的细菌;根据DGGE指纹图的聚类分析发现不同对虾及同一种对虾的鳃部与肠道内的细菌组成差异性非常大;同时也发现不同对虾体内有相同的细菌存在。首次尝试建立基于16S rDNA的PCR-DGGE基因指纹的对虾体内细菌组成揭示方法,对于今后建立对虾与养殖水体微生物和相关疾病的关系具有重要意义。     

Effect of cadmium exposure on lactate dehydrogenase activity in the hepatopancreas and abdominal muscle of the fiddler crab,Uca pugilator

1. Lactate dehydrogenase (LDH) activity in the hepatopancreas and abdominal muscle of fiddler crabs, Uca pugilator, was determined after 24 and 48 hr of exposure to 2 ppm cadmium chloride.2. For the cadmium exposed crabs, LDH activity in the hepatopancreas decreased, whereas that in the abdominal muscle increased.3. The increased LDH activity in the abdominal muscle may reflect increased dependence on anaerobic carbohydrate metabolism in fiddler crabs exposed to cadmium in their environment.     

In vivo humoral and cellular reactions, and fate of injected bacteria Aeromonas hydrophila in freshwater prawn Macrobrachium rosenbergii

Live non-opsonized and opsonized Aeromonas hydrophila were injected into juveniles of freshwater prawn Macrobrachium rosenbergii to study the cells involved in phagocytosis, distribution of bacteria, cellular reactions and clearance of both forms of bacteria from the system. The bacteria were rapidly distributed to various tissues viz., gills, heart, hepatopancreas within 1h, and the tissues revealed haemocytic nodule formation after 3 h of injection. There was rapid clearance of both the forms of bacteria from the circulation. However, clearance efficiency was significantly (P < 0.05) faster in the case of opsonized bacteria at 12 h after injection. Similarly, the nodule formation, that was prominent in cardiac musculature, was rapidly eliminated from the tissues of the group injected with opsonized bacteria as compared to non-opsonized bacteria injected group, thus confirming the existence of opsonic factors in haemolymph of this prawn. In another experiment, various dose levels of bacteria were injected intramuscularly into prawns and haemolymph was collected after 1, 6, 24, 72 h and 7 days of injection to study various immune parameters. Although, no major alterations in the total and differential haemocyte counts were observed in bacteria injected prawns compared to control, there was a significant decline in phenoloxidase activity in the highest dose bacteria injected group at the earlier phase and a rise in agglutinin levels at the later phase of the experimental period in the higher dose groups.     

Phosphamidon, methylparathion and dichlorvos impact on tissue oxidative metabolism in penaeid prawn, Metapenaeus monoceros

Changes in oxidative metabolism of hepatopancreas and muscle tissues of penaeid prawn, Metapenaeus monoceros was studied, following its exposure to selected organophosphorous insecticides phosphamidon, dichlorovos and methylparathion. The OPI are found to inhibit the activity levels of acetylcholinesterase, succinate dehydrogenase, isocitrate dehydrogenase, pyruvate dehydrogenase, lactate dehydrogenase and cytochrome-c-oxidase and cause accumulation of acetylcholine in the hepatopancreas and muscle tissues. These changes in the activity levels of selected oxidative enzymes during insecticide exposure in these tissues of prawn indicates the shift in the metabolic emphasis from aerobic to anaerobic conditions and is interpreted as a functional adaptation to insecticide induced metabolic stress. These observed changes at cellular level pave way for successful survival of prawns in insecticide polluted environ.     

Cloning and characterization of a hsp70 gene from Asiatic hard clam Meretrix meretrix which is involved in the immune response against bacterial infection

In the present study, a 71.43 kDa heat shock protein cDNA was cloned from Asiatic hard clam Meretrix meretrix. The cDNA was 2292 bp, containing an open reading frame (ORF) of 1959 bp, which encodes a protein of 652 amino acids with a theoretical molecular weight of 71.43 kDa and an isoelectric point of 5.32. Based on the amino acid sequence analysis and phylogenetic analysis, this hsp70 cDNA is a member of cytoplasmic hsc70 (constitutive genes) subfamily in the hsp70 family, and is designated as MmeHsc71. Quantitative RT-PCR was carried out to compare the spatial and temporal expression patterns of MmeHsc71 in the mRNA level between control clams and Vibrio parahaemolyticus-infected clams. Spatially, MmeHsc71 mRNA was found in all tested tissues, including foot, hepatopancreas, mantle and gill. MmeHsc71 mRNA expression level in hepatopancreas and gill displayed a significant increase in vibrio-challenged clams at 24h post-infection compared to control clams (P < 0.05). Temporally, there was a significant increase of MmeHsc71 mRNA level in hepathopancreas of vibrio-challenged clams compared to control clams at 6, 12, and 24h post-challenge, respectively. The result of quantitative immunofluorescence also indicated that there was obvious increase of MmeHsc71 in hepatopancreas of vibrio-challenged clams compared to control clams in protein level at 24h post-infection. The results suggested that MmeHsc71 may play an important role in mediating the immune responses of M. meretrix to bacterial challenge.