Tuula M. Aaltonen

Comparative effects of UVA and UVB irradiation on the immune system of fish.

Aquatic organisms can be harmed by the current levels of solar ultraviolet radiation. We have recently shown that exposure of fish to UVB irradiation alters the functioning of the fish immune system, but the effects of UVA radiation are unknown. The present study continues this work by characterizing UVA irradiation-induced immunological changes in fish. Roach, a cyprinid fish, were exposed to a single dose of either UVA (3.6 J/cm2) or UVB (0.5 J/cm2) irradiation. Both irradiations suppressed transiently mitogen-stimulated proliferation of blood lymphocytes. UVA, but not UVB, decreased hematocrit, plasma protein, and plasma immunoglobulin levels and increased the proportions of blood cells classified as unidentified leukocytes, possibly consisting of UVA-damaged lymphocytes. UVB, but not UVA, altered the functioning of head kidney and blood phagocytes, induced granulocytosis and lymphocytopenia in the blood and increased plasma cortisol concentration. These results imply that both UVA and UVB are potent modulators of the immune defence of fish.

Effects of ECF-Bleached Kraft Mill Effluent on Reproductive Steroids and Liver MFO Activity in Populations of Perch and Roach

Samples of perch (Perca fluviatilis L.) and roach (Rutilus rutilus L.) populations were collected in Lake Saimaa in September-October, after the onset of vitellogenesis and deposition of yolk in the eggs. The study area received elemental chlorine-free (ECF)-bleached kraft mill effluent (BKME). The pulp and paper mill studied used chlorine dioxide for bleaching and its effluents were treated in a modern activated sludge plant. The amounts of chlorophenolics in the bile of perch and roach at the mill site, 1 km from the discharge point, were two to four times higher than in fish from the upstream reference site. In the roach population near the mill liver the 7-ethoxyresorufin 0-deethylase (EROD) activity was four times that measured in the reference areas. However, the EROD activity in the perch remained unchanged. The liver 7-pentoxyresorufin 0-dealkylase (PROD) activity was induced to twice the amount only in male roach near the mill, whereas the relative liver size (LSI) was increased in female and male roach and female perch. The plasma concentrations of oestradiol-17β were decreased in female perch and female roach, but increased in male roach at the mill site. Near the mill, vitellogenin gene expression was also reduced in female perch. In addition, the plasma testosterone concentrations were reduced in female and male perch near the mill. No differences were noted in the plasma immunoglobulin concentrations in roach and in the relative gonad size (GSI) in the perch and roach populations. The condition factor was significantly lower only in female roach near the mill.

Ultraviolet B Irradiation Modulates the Immune System of Fish (Rutilus rutilus, Cyprinidae) II: Blood

Abstract The effects of a single dose of ultraviolet B (UVB) radiation (0.4 J/cm2) on immunological functions by blood leukocytes and on hematological parameters was studied in roach (Rutilus rutilus), a teleostean fish. The respiratory burst of phorbol 12-myristate 13-acetate stimulated whole blood phagocytes increased significantly after UVB irradiation but spontaneous cytotoxicity of blood leukocytes toward 51chromium-labeled K562 target cells was not markedly altered. Differential cell counting revealed that UVB exposure significantly increased the proportion of granulocytes and significantly decreased the proportion of lymphocytes in the peripheral blood, whereas hematocrit and the total number of white and red blood cells were unchanged. Plasma cortisol concentration increased in UVB-exposed fish. Severe handling stress caused similar, although not as potent, effects on the measured parameters of fish blood as UVB irradiation. These observations suggest that in fish UVB brings about a stress response, which may account for the observed alterations in the immune parameters and leukocyte composition of blood. Exposure of fish to strong visible light induced no alterations in immunological or hematological parameters, making it unlikely that ultraviolet radiation mediates its effects through visual perception.

Effects of primary- and secondary-treated bleached kraft mill effluents on the immune system and physiological parameters of roach.

The present study was designed to examine, whether, effluents from a modern pulp and paper mill using elemental chlorine-free/total chlorine-free (ECF/TCF) bleaching, exert effects on the immune system of fish and, in addition, to relate these findings to physiological parameters known to be affected by bleached kraft-mill effluents (BKME). Roach (Rutilus rutilus) were exposed in laboratory conditions to primary- or secondary-treated effluent from a pulp and paper mill. In order to study their capability to respond to foreign antigens they were immunised with bovine gamma-globulin (BGG) prior to exposure. The number of anti-BGG antibody-secreting cells (ASC) and the number of immunoglobulin-secreting cells (ISC) in the spleen and blood as well as the level of anti-BGG specific antibodies and concentration of plasma immunoglobulin (IgM) were studied. Phagocytosis and migration of granulocytes of the head kidney were also determined. In addition to the immunological parameters, the activity of hepatic biotransformation enzymes, the carbohydrate metabolism and osmoregulation were examined. Exposure of roach for 21 days to BKME affected several immunological parameters. Both effluents, primary- and secondary-treated, impaired the immunoreactivity of the fish. Sex-related differences in the immune responses were evident in many parameters e.g. in the number of blood ISC and splenic ASC. Sex also had effects on cortisol levels and in the induction of 7-ethoxyresorufin O-deethylase (EROD). These results demonstrate that both primary- and secondary-treated effluent from a pulp and paper mill using ECF/TCF bleaching have effects on fish immune functions. Further, these findings suggest that steroids may contribute to immunomodulation in fish.

Ultraviolet B Irradiation Modulates the Immune System of Fish (Rutilus rutilus, Cyprinidae) Part III: Lymphocytes¶

Abstract The effects of short-term exposure to ultraviolet B (UVB) radiation on lymphocyte-related parameters were studied under controlled laboratory conditions using roach (Rutilus rutilus), a cyprinid teleost, as the model fish. In vitro lymphoproliferative responses stimulated with a T-cell–specific mitogen, concanavalin A (ConA), or a B-cell–specific activator, lipopolysaccharide (LPS), were decreased in exposed fish. Also nonstimulated proliferation was lower than in unexposed fish. ConA-activated responses returned to normal levels within 7 days after exposure, but LPS-activated responses were reduced throughout the 14 day follow-up. The capability of UVB-exposed fish to produce an antibody response was studied by intraperitoneal immunization with bovine γ-globulin (BGG). The concentration of anti-BGG antibodies in plasma as well as the number of anti-BGG–specific antibody-secreting cells in the spleen or blood were not decreased in fish exposed either to a single dose of UVB prior to immunization, or to single dose of UVB prior to immunization followed by three additional doses after immunization. Immunoglobulin M (IgM) production, when assayed as plasma IgM level or as the number of IgM-secreting cells in the spleen or blood, was not suppressed after exposure to UVB irradiation. These results indicate that a single dose of UVB or short-term exposure to UVB irradiation has no negative effects on IgM production or reactivity against antigen administered via the intraperitoneal route. However, the suppression of in vitro lymphoproliferative responses suggest that exposure to UVB has the potential to interfere with lymphocyte-related functions in fish.

Effects of ultraviolet light on immune parameters of the roach

Ultraviolet B radiation penetrates into water and can affect fish health and the immune system, as is the case with mammals. Teleost fish, the roach, were exposed to UVB irradiation in aquariums and a panel of immune parameters was determined. In addition to altered blood picture and respiratory burst by blood leukocytes, changes were noted also in major lymphatic organs. Respiratory burst and natural cytotoxicity activity of head kidney granulocytes and mitogen-activated proliferation of splenic lymphocytes were suppressed. Although mostly transitory, some parameters remained suppressed for the following 2 weeks. Ultraviolet A radiation had only minor effects. The stress induced by UVB may be involved in the modulation of immune parameters.

Modulation of immune parameters of roach, Rutilus rutilus, exposed to untreated ECF and TCF bleached pulp effluents

Abstract The present study was designed to assess the effects of elemental chlorine free (ECF) and totally chlorine free (TCF) pulp and paper mill effluents on the immune defence of the roach ( Rutilus rutilus ). Fish were exposed for 5 weeks to concentrations of 0, 0.6, 2, 6 and 20% of untreated effluents from ECF or TCF bleaching processes. In order to study the capability to respond to foreign antigens the fish were immunised with bovine γ-globulin (BGG) 3 weeks before sampling. The numbers of anti-BGG antibody-secreting cells and immunoglobulin-secreting cells in the spleen and blood, as well as the levels of the anti-BGG antibodies and concentrations of immunoglobulin in plasma and the proliferation of lymphocytes after mitogenic activation were determined. In addition, phagocytosis-related functions, respiratory burst activity and migration of leukocytes, were measured. Formation of specific anti-BGG antibody-secreting cells in the blood and immunoglobulin-secreting cells in the spleen and blood were suppressed in the fish exposed to a high concentration (20%) of ECF effluent. Similarly, the level of anti-BGG antibody and the stimulation indices of Concanavalin A-stimulated proliferation responses in vitro were lower in these fish. A decreased migration of granulocytes was observed in the fish exposed to 0.6–2% of ECF and TCF effluents. These results demonstrate that untreated ECF and TCF effluents significantly affected the immune parameters of the roach. Taken as a whole, fish exposed to TCF effluent showed less immunomodulation than those exposed to EFC effluent.

Experimental infection of brown trout, Salmo trutta L., using a Finnish Yersinia ruckeri isolate

Abstract The pathogenicity of a Finnish Yersinia ruckeri isolate to 1-year-old brown trout was tested using both immersion and injection mode of infection. The occurrence of bacteria and antibody production were monitored for up to 25 days. No disease signs or mortality due to Y. ruckeri occurred in the immersion experiment where the fish were further stressed with Ichthyophthirius multifiliis , but Y. ruckeri was isolated from certain organs in half of the fish. The intestine, kidney and spleen were the most favoured sites of infection, only one fish having bacteria in the liver. The injection of brown trout caused 100% Y. ruckeri infection and some mortality was recorded during the first week post infection. All dead fish exhibited signs of bacterial infection but none had inflammation around the mouth. Antibodies against Y. ruckeri were produced in both the groups subjected to immersion and that subjected to injection but the production was higher in the injected fish. These results confirm earlier findings that the Finnish Y. ruckeri is less pathogenic than, for example, Y. ruckeri in USA, and indicate that more experimental studies are needed to examine the significance of geographical variation of Y. ruckeri strains in terms of their pathogenicity.