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  • 1.
    Hernroth, Bodil
    The Royal Swedish Academy of Sciences.
    Factors influencing bactericidal activity of blue mussel (Mytilus edulis) haemocytes against Salmonella typhimurium2003In: Fish and Shellfish Immunology, ISSN 1050-4648, E-ISSN 1095-9947, Vol. 14, no 2, p. 93-104Article in journal (Refereed)
    Abstract [en]

    This study showed that in vitro survival of Salmonella typhimurium, after exposure to haemocytes of Mytilus edulis, was significantly affected by the lipopolysaccharides (LPS) structures expressed on the cell surface of the bacteria. Survival seemed to be affected by the surrounding temperature as well. Mussel haemocytes were in vitro exposed to mutants of S. typhimurium, expressing differences in O -antigen polysaccharide chains and core sugars of LPS on their cell surface. Surviving cells of the mutants were determined after incubation with the haemocytes at different temperatures, using a colorimetric assay. In addition, a complementary study on clearance of these mutants, inoculated into the adductor muscle of mussels, was performed at 6 and 20 degrees C. It was concluded that the survival index (SI%) measured in vitro for the mutant with complete LPS was significantly lower at 6 degrees C (c.15%) compared to that at 14 and 20 degrees C (c.70%). SI% for the other mutants was c.35-45% and was not affected by temperature. The in vivo study at 20 degrees C showed that during the first 24h, the clearance rate for the mutants with complete LPS was significantly higher than for the others. Thereafter all mutants, with exception for the most deficient, started to increase in numbers and caused death to the mussels. At 6 degrees C the mutants were slowly reduced and after 17 days, viable cells of the mutant with complete LPS were still detectable in the haemolymph. The study indicated that the mussel haemocytes responded in relation to the LPS of the mutants. However, more intact LPS also seemed to protect the bacteria from being killed. The higher temperatures favoured the growth of the mutants that managed to resist the haemocyte defence. Cell surface properties and temperature seem to affect the survival of bacteria in mussels, which consequently can affect risk assessments in regard to public health.

  • 2.
    Hernroth, Bodil
    The Royal Swedish Academy of Sciences.
    The influence of temperature and dose on antibacterial peptide response against lipopolysaccharide in the blue mussel, Mytilus edulis.2003In: Fish and Shellfish Immunology, ISSN 1050-4648, E-ISSN 1095-9947, Vol. 14, no 1, p. 25-37Article in journal (Refereed)
    Abstract [en]

    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.

  • 3.
    Hernroth, Bodil
    et al.
    Kristianstad University, School of Education and Environment, Avdelningen för Naturvetenskap. Kristianstad University, Research environment Man & Biosphere Health (MABH).
    Baden, S.
    University of Gothenburg.
    Tassidis, Helena
    Kristianstad University, School of Education and Environment, Avdelningen för Naturvetenskap. Kristianstad University, Research environment Man & Biosphere Health (MABH).
    Hörnaeus, K.
    Uppsala University.
    Guillemant, J.
    Uppsala University.
    Bergström Lind, S.
    Uppsala University.
    Bergquist, J.
    Uppsala University.
    Impact of ocean acidification on antimicrobial activity in gills of the blue mussel (Mytilus edulis)2016In: Fish and Shellfish Immunology, ISSN 1050-4648, E-ISSN 1095-9947, Vol. 55, p. 452-459Article in journal (Refereed)
    Abstract [en]

    Here, we aimed to investigate potential effects of ocean acidification on antimicrobial peptide (AMP) activity in the gills of Mytilus edulis, as gills are directly facing seawater and the changing pH (predicted to be reduced from ∼8.1 to ∼7.7 by 2100). The AMP activity of gill and haemocyte extracts was compared at pH 6.0, 7.7 and 8.1, with a radial diffusion assay against Escherichia coli. The activity of the gill extracts was not affected by pH, while it was significantly reduced with increasing pH in the haemocyte extracts. Gill extracts were also tested against different species of Vibrio (V. parahaemolyticus, V. tubiashii, V. splendidus, V. alginolyticus) at pH 7.7 and 8.1. The metabolic activity of the bacteria decreased by ∼65–90%, depending on species of bacteria, but was, as in the radial diffusion assay, not affected by pH. The results indicated that AMPs from gills are efficient in a broad pH-range. However, when mussels were pre-exposed for pH 7.7 for four month the gill extracts presented significantly lower inhibit of bacterial growth. A full in-depth proteome investigation of gill extracts, using LC-Orbitrap MS/MS technique, showed that among previously described AMPs from haemocytes of Mytilus, myticin A was found up-regulated in response to lipopolysaccharide, 3 h post injection. Sporadic occurrence of other immune related peptides/proteins also pointed to a rapid response (0.5–3 h p.i.). Altogether, our results indicate that the gills of blue mussels constitute an important first line defence adapted to act at the pH of seawater. The antimicrobial activity of the gills is however modulated when mussels are under the pressure of ocean acidification, which may give future advantages for invading pathogens.

  • 4.
    Hernroth, Bodil
    et al.
    Kristianstad University, Faculty of Natural Science, Research environment Man & Biosphere Health (MABH). KVA.
    Baden, S
    Gothenburg University.
    Tassidis, Helena
    Kristianstad University, Faculty of Natural Science, Research environment Man & Biosphere Health (MABH). Kristianstad University, Faculty of Natural Science, Avdelningen för miljö- och biovetenskap.
    Hörnaeus, K
    Uppsala University.
    Guillemant, J
    Uppsala University.
    Bergström Lind, S
    Uppsala University.
    Bergquist, J
    Uppsala University.
    Impact of oceanacidification on antimicrobial activity in gills of the blue mussel (Mytilusedulis)2016In: Fish and Shellfish Immunology, ISSN 1050-4648, E-ISSN 1095-9947, Vol. 55, p. 452-459Article in journal (Refereed)
  • 5.
    Hernroth, Bodil
    et al.
    Kristianstad University, School of Education and Environment.
    Nilsson Sköld, Helen
    Department of Biological and Environmental Sciences, Kristineberg, University of Gothenburg.
    Wiklander, Kerstin
    Mathematical Sciences, University of Gothenburg.
    Jutfelt, Fredrik
    Department of Biological and Environmental Sciences, University of Gothenburg.
    Baden, Susanne
    Department of Biological and Environmental Sciences, Kristineberg, University of Gothenburg.
    Simulated climate change causes immune suppression and protein damage in the crustacean Nephrops norvegicus2012In: Fish and Shellfish Immunology, ISSN 1050-4648, E-ISSN 1095-9947, Vol. 33, no 5, p. 1095-1101Article in journal (Refereed)
    Abstract [en]

    Rising atmospheric carbon dioxide concentration is causing global warming, which affects oceans by elevating water temperature and reducing pH. Crustaceans have been considered tolerant to ocean acidification because of their retained capacity to calcify during subnormal pH. However, we report here that significant immune suppression of the Norway lobster, Nephrops norvegicus, occurs after a 4-month exposure to ocean acidification (OA) at a level predicted for the year 2100 (hypercapnic seawater with a pH lowered by 0.4 units). Experiments carried out at different temperatures (5, 10, 12, 14, 16, and 18 °C) demonstrated that the temperature within this range alone did not affect lobster immune responses. In the OA-treatment, hemocyte numbers were reduced by almost 50% and the phagocytic capacity of the remaining hemocytes was inhibited by 60%. The reduction in hemocyte numbers was not due to increased apoptosis in hematopoetic tissue. Cellular responses to stress were investigated through evaluating advanced glycation end products (AGE) and lipid oxidation in lobster hepatopancreata, and OA-treatment was shown to significantly increase AGEs', indicating stress-induced protein alterations. Furthermore, the extracellular pH of lobster hemolymph was reduced by approximately 0.2 units in the OA-treatment group, indicating either limited pH compensation or buffering capacity. The negative effects of OA-treatment on the nephropidae immune response and tissue homeostasis were more pronounced at higher temperatures (12–18 °C versus 5 °C), which may potentially affect disease severity and spread. Our results signify that ocean acidification may have adverse effects on the physiology of lobsters, which previously had been overlooked in studies of basic parameters such as lobster growth or calcification.

  • 6.
    Oweson, Carolina
    et al.
    Institute of Marine Ecology, University of Gothenburg.
    Hernroth, Bodil
    Kristianstad University College, School of Teacher Education.
    A comparative study on the influence of manganese on the bactericidal response of marine invertebrates2009In: Fish and Shellfish Immunology, ISSN 1050-4648, E-ISSN 1095-9947, Vol. 27, no 3, p. 500-507Article in journal (Refereed)
    Abstract [en]

    Manganese, Mn, is a naturally abundant metal in marine sediments. During hypoxic conditions the metal converts into a bioavailable state and can reach levels that have been shown immunotoxic to the crustacean Nephrops norvegicus. For this species it has previously been shown that exposure to 15 mg L(-1) of Mn decreased the number of circulating haemocytes while it for the echinoderm Asterias rubens increased the number of coelomocytes. Here, we compared if five days of exposure to the same concentration of Mn affects the bactericidal capacity of these two species and the mollusc Mytilus edulis when inoculated with the bacterium Vibrio parahaemolyticus. Viable counts of the bacteria were investigated at a time-course post-injection in the blood and the digestive glands of Mn-exposed and un-exposed (controls) animals. Accumulation of Mn was also analyzed in these tissues. When exposed to Mn the haemocyte numbers were significantly reduced in M. edulis and it was shown that the bactericidal capacity was impaired in the mussels as well as in N. norvegicus. This was most obvious in the digestive glands. These two species also showed the highest accumulation of the metal. In A. rubens the bactericidal capacity was not affected and the metal concentration was similar to the exposure concentration. After a recovery period of three days the concentration of Mn was significantly reduced in all three species. However, in M. edulis and N. norvegicus it was still double that of A. rubens which could explain the remaining bactericidal suppression observed in N. norvegicus. This study pointed out that exposure to such Mn-levels that are realistic to find in nature could have effects on the whole organism level, in terms of susceptibility to infections. The effect seemed associated to the accumulated concentration of Mn which differed on species level.

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