Neuroendocrine-immune interactions in fish: a role for interleukin-1

Bi-directional communication between the hypothalamus-pituitary-adrenal (HPA)-axis and the sympathetic nervous system with the immune system is crucial to ensure homeostasis. Shared use of ligands and especially receptors forms a key component of this bi-directional interaction. Glucocorticoids (GC), the major end products of the HPA-axis differentially modulate immune function. Cytokines, especially interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), ensure immune signalling to the neuroendocrine system. In addition, hormones from leukocyte origin such as corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and beta-endorphin, as well as centrally synthesised and secreted cytokines, contribute to the communication network.In teleost fish cortisol is the major product of the hypothalamus-pituitary-interrenal (HPI)-axis which is the teleost equivalent of the HPA-axis. Moderate and substantial increases in cortisol during stressful circumstances negatively affect B-lymphocytes, whereas rescue of neutrophilic granulocytes may support innate immunity. Recent elucidation of lower vertebrate cytokine sequences has facilitated research into neuroendocrine-immune interactions in teleosts and the first evidence for a significant function of interleukin-1 in the bi-directional communication is discussed.     

Disease resistance and growth of two inbred carp Cyprinus carpio L. lines and their hybrid in pond culture

Recently, we examined the specific resistance of two carp lines (W and R8) and their hybrid, to experimental bath challenges with atypical Aeromonas salmonicida. These experiments indicated a genetic difference in resistance, the W carp being most susceptible. In the present study, fish of the same spawnings were grown in pond culture, to examine the correlation between disease resistance observed in the laboratory and survival rate in the field, as well as to examine the production potential of the hybrid. Hybrid advantage resulted in higher average weights at the field samplings. Correlation of the field survival rates with specific resistance to A. salmonicida was pronounced. Survival of the W carp after winter was much lower (47%) than survival of the R8 (93%) and hybrid carp (93%).     

The immune system of cyprinid fish. Oxytetracycline and the regulation of humoral immunity in carp (Cyprinus carpio)

The antibiotic oxytetracycline (oxyTC) was administered either by mixing with food or by intraperitoneal injections. In oxyTC treated animals decreased serum immunoglobulin levels were found. The primary anti-sheep red blood cell (SRBC) response was measured by enumerating plaque forming cells (PFC). It was observed that the PFC response was depressed by 80–95% in oxyTC treated animals. When an anti-SRBC serum was injected together with SRBC the immunosuppressive effect of oxyTC was absent. A secondary anti-SRBC response was not inhibited by oxyTC. On base of the results a model for antigen presentation and the interaction between macrophages, T- and B-like cells during primary and secondary responses in fish is proposed. In primary responses cellular interaction is needed to develop a proper immune response whereas after a high antigen dose challenge SRBC might behave as a T-independent antigen.     

The influence of antibiotics on the immune system. II. Modulation of fish leukocyte responses in culture

We report the immunomodulating effect of two tetracycline analogues (oxytetracycline and doxycycline) on mitogenic and allogeneic stimulation of carp (Cyprinus carpio) leukocytes in vitro. Both drugs interfered with 3H-thymidine incorporation into the DNA of phytohaemagglutinin-stimulated pronephric leukocytes in a dose-dependent manner. Low concentrations of oxytetracycline delayed the mitogenic response, but did not reduce it. In addition, several other antibiotics were tested for their possible interference with blastogenesis.     

The use of artificial gynogenesis in studies on the immune system of carp (Cyprinus carpio L.)

Gynogenetic carp were obtained by using gamma-irradiated sperm for fertilization followed by a cold shock. The gynogenetic fish and normal siblings of the same parents were analysed for 3 genetic markers: scale pattern, transferrin (Tf) and allograft rejection. Gynogenetic offspring showed only the maternal scale pattern (scattered). In the majority of the gynogenetic fish a heterozygous Tf pattern identical to the mother was observed. Animals homozygous for Tf were rare in this group. A typical paternal Tf band was only seen in normal siblings. An exchange of skin grafts between normal animals was always followed by rejection (median survival time, 12 days at 23 OC). However, the rejection was delayed in the gynogenetic group (MST, 19,6 days). There were even signs of acceptance in some animals. This approach can be used for the identification of 2nd and 3rd generation gynogenetic fish, which are homozygous for certain MHC haplotypes.     

Enzymatic production and characterization of konjac glucomannan oligosaccharides

Enzymes from a balanced human gut flora are promising tools to design prebiotic oligosaccharides. In this study, we investigated the action of enzymes from fecal bacteria on the complex polysaccharide konjac glucomannan (KGM). The oligosaccharides produced were compared to oligosaccharides from KGM digests with fungal endo-β-(1,4)-glucanase (EG) or endo-β-(1,4)-mannanase (EM). For this purpose, the oligosaccharides from the different digests were first studied for their structural characteristics like monosugar composition and exo-enzymatic degradability, as monitored by capillary electrophoresis with laser-induced fluorescence detection. Whereas the oligosaccharides produced by EG and EM were characteristic for the selectivity of the respective enzyme in cleaving the mannose-/glucose-sugar linkages of KGM, oligosaccharides produced by the fecal enzymes did not point to a sugar-selective degradation. The oligosaccharide fragments from the different digests indicated the KGM polysaccharide to be composed of a backbone composed of short mannose and glucose sequences, to which branches rich in mannose are attached.     

Rectal tears in the horse: malpractice or an unfortunate accident?

Rectal tears are a relatively rare complication of rectal palpation, mating or dystocia, and idiopathic spontaneous occurrence of rectal perforation has also been described. Rectal tears have been classified in a three or a four-grade system. Immediate recognition of the fact that a tear has been made and prompt action will improve the horses chance of survival, and is the best defence of the veterinarian against legislation. There is no clear explanation why rectal tears sometimes occur. A questionnaire of the Netherlands Equine Veterinary Association revealed that rectal tears occurred both to inexperienced veterinarians and to very experienced colleagues, working with or without a probe for ultrasonography. In the opinion of the authors, the approach in the legislation of rectal tears should be that a rectal tear is only considered 'malpractice' if the veterinarian obviously acted carelessly. However, this would create the ridiculous situation that it seems better for the veterinarian-client relationship when the veterinarian confesses to 'careless' rectal palpation. This point needs further clarification, and perhaps an 'insurance of the horse for rectal palpation' will be the solution. Further, retrospectively over the last ten years, the problem is often not 'making a rectal tear', but 'lacking recognition of making a tear'. Whenever a rectal tear is suspected, the veterinarian must assess its severity, discuss the problem openly with the owner and apply appropriate treatment including referral.     

The immune system of cyprinid fish. The immunosuppressive effect of the antibiotic oxytetracycline in carp (Cyprinus carpio L.)

The effect of oxytetracycline (oxyTC) upon the immune system of carp was investigated. OxyTC was administered by feeding with oxyTC-containing pellets or by intraperitoneal injection. In order to study cellular immunity, allogeneic scale transplantation was carried out. Oral administration of oxyTC had no influence upon the median survival time (MST) of the scales. However, injections with oxyTC significantly prolonged the MST from 8.5 to 11–20 days. Thus, cellular immunity was not affected by oral administration of oxyTC, but injections did have a dramatic immunosuppressive effect.To investigate the effect of oxyTC upon humoral immunity, animals were injected with rabbit red blood cells (RaRBC). During the primary and secondary response the number of rosette forming cells (RFC) in the spleen was determined. Control animals (not treated with oxyTC) developed an anti-RaRBC response up to 25000 RFC/10⁶ white spleen cells but oxyTC-treated animals always showed reduced RFC numbers. In some cases the RFC number in oxyTC-treated animals was comparable with background levels in non-immunized control animals (4000 RFC/10⁶ white spleen cells). Thus, irrespective of the route of administration, the humoral immune response is depressed by oxyTC. It is concluded that both humoral and cellular immune responses of carp are suppressed during treatment with oxyTC.Preliminary observations showed an increased number of granulocytes in the spleen of oxyTC-treated animals. It is tempting to speculate that in those cases where specific lymphoid defence mechanisms are blocked, the phagocytic defence system becomes more active.     

Blood changes in carp (Cyprinus carpio) induced by ulcerative Aeromonas salmonicida infections

Only recently Aeromonas salmonicida has been recognized as a significant bacterial pathogen in ulcerative disease of cyprinid fish. Our attempts to formulate a vaccine based on bacterial surface antigens were unsuccessful in conferring reliable protection against lethal challenge. This lead us to study pathological changes in the humoral defense system during ulcerative A. salmonicida infection in carp. High numbers of opportunist pathogens such as A. hydrophila and Pseudomonas sp. were frequently recovered from the internal organs of moribund fish, in addition to A. salmonicida. These findings together with leucopenia in moribund fish suggest that pathogenesis is characterized by a state of immune suppression. In addition, fish which had sustained a sublethal infection were not protected against a subsequent lethal challenge. However, fish previously injected with a concentrated and inactivated culture supernatant showed protection. Differential blood cell counts did not differ between experimental and control groups during sublethal infection in contrast to serum proteins. Furthermore infected non-immune carp showed a progressive decrease of immunoglobulin and total serum protein levels before the day of peak mortality whereas protected carp maintained the immunoglobulin concentration despite a decrease in protein. Our observations suggest the involvement of multiple pathogenic events, affecting different parts of the humoral defense system during ulcerative A. salmonicida infection. The immunosuppressive effects can be minimized by prior vaccination with culture supernatant.