Seasonal changes in hemocytes and serum defense factors in the blue mussel Mytilus galloprovincialis

Santarem, Marco; Robledo, J.A.F.; Figueras, Antonio

doi:10.3354/dao018217

Cited by 76 publications

(39 citation statements)

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“…This variation may reflect the biological characteristics of the individuals, their physiological state and the environmental conditions (Auffret & Oubella 1995), and may be related to the haemopoietic activity of the bivalves (Anderson 1990, Pipe et al 1995. The number of haemocytes were influenced by seasonal factors (such as temperature, reproductive cycle, food), as reported by Santarèm et al (1994) for Mytilus galloprovincialis. Feng (1965) reported that the number of circulating haemocytes in Crassostrea virginica is influenced by the amount of food and the temperature.…”

Section: Discussionmentioning

confidence: 94%

Morphological and cytoenzymatic characterization of haemocytes of the venus clam Chamelea gallina

Pampanin

Marin²,

Ballarin³

2002

Dis. Aquat. Org.

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A morphological and enzymatic characterization of Chamelea gallina haemocytes was carried out as a prerequisite for further studies on venus clam immunobiology. Two main types of circulating haemocytes were identified (1) hyalinocytes (79.2%), agranular cells with a central nucleus surrounded by a little cytoplasm, and (2) granulocytes (16.5%), smaller granular cells with smaller nuclei. Small cells with a strongly basophilic nucleus and a thin layer of peripheral cytoplasm, probably undifferentiated blast cells (4.3%), were also observed. Both granulocytes and hyalinocytes can assume a spreading or round morphology. The enzymatic activities of haemocytes were also investigated. Some of the granulocytes and hyalinocytes were positive for hydrolytic enzymes, suggesting a role for these cells in phagocytosis; no oxidative enzymes were detected in C. gallina haemocytes. Granulocytes and hyalinocytes can easily adhere to the substratum and exhibit a low phagocytosis activity towards foreign particles (6.3%), whereas the fraction of cells containing ingested material significantly increased after pre-incubation of test particles with cell-free haemolymph, which suggests the presence of opsonin(s) in the haemolymph. KEY WORDS: Haemocytes · Chamelea gallina · Cytoenzymology · Phagocytosis Resale or republication not permitted without written consent of the publisherDis Aquat Org 49: [227][228][229][230][231][232][233][234] 2002 (Cheney 1971, Foley & Cheng 1974, Moore & Eble 1977, Huffman & Tripp 1982. Since no data are available concerning Chamelea gallina haemocytes and their role in immunity, we carried out a morpho-functional characterization of C. gallina haemocytes as a prerequisite for further studies on the immunobiology of the venus clam in order to define a series of immuno-biomarkers to be used in the evaluation of stress conditions for this species.Our results indicate the presence of 2 main haemocyte types, namely granulocytes and hyalinocytes, both exhibiting lysosomal enzymes and phagocytosis activity. MATERIALS AND METHODSClams (shell length 20 to 30 mm) were collected with a hydraulic dredge at Venice Lido, on the west coast of the Northern Adriatic Sea, in front of the Venice lagoon. They were kept in the laboratory for 2 to 3 d before experiments in 70 l tanks filled with aerated seawater that was renewed every other day (salinity 35 ± 1 ‰; temperature 17 ± 1°C) and fed with microalgae (Isochrysis galbana and Phaeodactylum tricornutum).Heamolymph collection. Haemolymph was collected from the anterior adductor muscle of clams with a syringe containing 10 mM of L-cysteine in filtered seawater (FSW), pH 7.5, to a final dilution of 1:1 haemolymph/L-cysteine. Haemolymph was also collected in the absence of L-cysteine and centrifuged at 780 × g for 10 min to pellet cells and obtain cell-free haemolymph (CFH).Haemocyte cultures. Haemolymph was centrifuged at 780 × g for 10 min and the pellet was resuspended in FSW to achieve a final concentration of 10 6 cells ml -1 . Sixty µl of haemocyte susp...

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Section: Discussionmentioning

confidence: 94%

Morphological and cytoenzymatic characterization of haemocytes of the venus clam Chamelea gallina

Pampanin

Marin²,

Ballarin³

2002

Dis. Aquat. Org.

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“…A possible linking factor between these latter elements could be the reproductive cycle, which can affect haemocytes and which is also a means by which molluscs can remove hydrocarbons. A field study in Spain on Mytilus galloprovincialis recorded strong seasonal effects on haemocyte numbers, which were attributed principally to reproductive effects (Santarém et al 1994). Alternatively, factors such as salinity or temperature can influence bivalve immunity (Fisher & Newall 1986, Chu & LaPeyre 1993, although they may have less direct roles than the reproductive cycle in hydrocarbon accumulation/depuration.…”

Section: Discussionmentioning

confidence: 99%

Changes in immune parameters of natural mussel Mytilus edulis populations following a major oil spill ('Sea Empress', Wales, UK)

Dyrynda¹,

Law²,

Dyrynda³

et al. 2000

Mar. Ecol. Prog. Ser.

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In February 1996, the oil tanker 'Sea Empress' spilt over 70 000 t of crude oil which contaminated ca 200 km of coastline (Milford Haven, Wales, UK). The effects of the oil on immunity in mussels Mytilus edulis were investigated in parallel with the measurement of hydrocarbon contamination in the tissues. Initially, severe immunosuppression occurred in oiled mussels, corresponding with very high polycyclic aromatic hydrocarbon (PAH) levels. The haemocytes of mussels from oiled sites showed significantly reduced superoxide generation and phagocytic activity, effects likely to have deleterious consequences for successful disease resistance. As contaminant levels decreased, the immunosuppression became less extreme and recovery was evident by May 1996. Between October 1996 and March 1997, immune activity in the haemocytes of the previously oiled mussels was again significantly reduced, coinciding with increased PAH levels. During this latter period, certain high molecular mass PAHs (characteristically derived from combustion processes) were primarily responsible for the increase, occurring at similar concentrations in the mussel tissues to those observed just after the spill. A subsequent reduction of hydrocarbons in June 1997 was followed by another, but less marked, increase in PAHs between October 1997 and March 1998, coupled with only minimal changes in immunity. The results show that immunosuppression following the oil spill was severe, but that recovery followed a few months later and the initial effects were not therefore permanent. The results also suggest that seasonal peaks in combustion-derived PAHs may occur in the region and that these would have been greatly exacerbated early in 1996 by oil released from the 'Sea Empress'. KEY WORDS: Mytilus · Immune defence · Haemocyte · Oil spill · Hydrocarbon contaminationResale or republication not permitted without written consent of the publisher Mar Ecol Prog Ser 206: 155-170, 2000 These include the induction of biomarker enzymes in fish and molluscs (Sole et al. 1996, Woodin et al. 1997, Stagg et al. 1998, Kirby et al. 1999, abnormalities in fish embryos/larvae (Hose et al. 1996, Bue et al. 1998 and haematological changes in vertebrates (Duffy et al. 1994, Walton et al. 1997.Despite the considerable effects that crude oil contaminants can exert on the physiology of marine animals, information concerning impacts on the physiology of natural invertebrate populations is more limited, particularly with respect to their immune defences. Globally, marine invertebrates constitute an important economic resource, and factors influencing their ability to resist disease and hence maintain healthy populations are of considerable relevance. Invertebrate immune defences comprise cellular and humoral components, which are highly efficient in combating pathogens and parasites and share many facets with vertebrate immune defences (Ratcliffe et al. 1985, Engstrøm et al. 1993. Laboratory exposure to contaminants can significantly alter immunity in invertebrates fr...

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“…Lysozyme activity was quantified according to Santarem et al (1994). CL (50 µl), obtained as described above, was added to 950 µl of a 0.15% suspension of Micrococcus lysodeikticus (Sigma) in 66 mM phosphate buffer, pH 6.2, and the decrease in absorbance (∆A min -1 ) was continuously recorded at 450 nm for 5 min at 20°C.…”

Section: Animalsmentioning

confidence: 99%

4-Nonylphenol induces immunomodulation and apoptotic events in the clam Tapes philippinarum

Matozzo¹,

Marin

2005

Mar. Ecol. Prog. Ser.

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The effects of 4-nonylphenol (NP) on functional responses of haemocytes from the clamTapes philippinarum were investigated after 7 d exposure to sublethal NP concentrations (0, 0 + acetone, 0.025, 0.05, 0.1 and 0.2 mg l -1 NP). Haemocytes from both controls and exposed clams were collected, and the effects of NP on uptake of the vital dye Neutral Red (NR), superoxide dismutase (SOD) and lysozyme activities, total haemocyte count (THC) and volume of circulating cells were evaluated. The capability of NP to induce apoptosis was also investigated. Exposure of clams to 0.2 mg l -1 NP significantly increased (p < 0.05) NR uptake when compared with controls, suggesting that NP caused alterations in cell membrane stability. Significant decreases in both SOD and lysozyme activity were observed from 0.05 mg l -1 NP (p < 0.01 and p < 0.05, respectively) with respect to controls, indicating that NP causes oxidative stress and reduces the immunocompetence of the exposed clams. From 0.05 mg l -1 NP, the apoptotic index (percentage of haemocytes showing positivity to the TUNEL reaction) significantly increased (p < 0.001). Apoptotic haemocytes showed shrinkage and lost their amoeboid shape. Moreover, NP exposure significantly increased THC (p < 0.05 at 0.2 mg l -1 NP) and caused a different distribution in size frequency. On the basis of the threshold concentration values, the most sensitive endpoints for NP were SOD, lysozyme and apoptosis assays. Our results highlight a relationship between NP exposure and changes in the functional responses of haemocytes, suggesting that the contaminant induces immunomodulation in T. philippinarum, mainly by altering cell morphology, cell membrane stability and enzymatic activities, and by promoting oxidative stress and apoptotic events.

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Seasonal changes in hemocytes and serum defense factors in the blue mussel Mytilus galloprovincialis

Cited by 76 publications

References 20 publications

Morphological and cytoenzymatic characterization of haemocytes of the venus clam Chamelea gallina

Morphological and cytoenzymatic characterization of haemocytes of the venus clam Chamelea gallina

Changes in immune parameters of natural mussel Mytilus edulis populations following a major oil spill ('Sea Empress', Wales, UK)

4-Nonylphenol induces immunomodulation and apoptotic events in the clam Tapes philippinarum

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