Influence of chronic stress on the mechanism of the cytotoxic system in common carp (Cyprinus carpio).

Aquaculture conditions expose fish to internal and environmental stressors that increase their susceptibility to morbidity and mortality. The brain accumulates stress signals and processes them according to the intensity, frequency duration and type of stress, recruiting several brain functions to activate the autonomic or limbic system. Triggering the autonomic system causes the rapid release of catecholamines, such as adrenaline and noradrenaline, into circulation from chromaffin cells in the head kidney.

Acute stress promotes post-injury brain regeneration in fish.

The central nervous system and the immune system, the two major players in homeostasis, operate in the ongoing bidirectional interaction. Stress is the third player that exerts strong effect on these two 'supersystems'; yet, its impact is studied much less. In this work employing carp model, we studied the influence of preliminary stress on neural and immune networks involved in post-injury brain regeneration.

The protective effect of humic-rich substances on atypical Aeromonas salmonicida subsp. salmonicida infection in common carp (Cyprinus carpio L.).

When challenged with atypical Aeromonas salmonicida subsp. salmonicida, exposure of the common carp (Cyprinus carpio L.) to different humic-rich compounds resulted in a significant reduction in infection rates.

A deleterious effect associated with UNH159 is attenuated in twin embryos of an inbred line of blue tilapia Oreochromis aureus.

Offspring of a highly inbred gynogenetic line of Oreochromis aureus displayed 12-fold increase in twinning rate compared to the outbred population. Asymmetric conjoined twins, which consist of a normal embryo attached to a malformed-atrophic twin, were frequently encountered in both gynogenetic (90·7%) and outbred (38·2%) embryos. The monozygotic origin of these twins was determined using five microsatellite markers.

Application of ultrasound in vaccination against goldfish ulcer disease: a pilot study.

Antigenicity and immunogenicity of Aeromonas salmonicida soluble A-layer protein (AP) and formalin-killed bacterin were assessed in goldfish (Carassius auratus L.) by applying ultrasound-immersion (USI) and injection (Inj) routes. No adjuvant was used in USI while IFA was used in Inj.

Vertical and horizontal transmission of tilapia larvae encephalitis virus: the bad and the ugly.

Impairment of innate immunity in tilapia larvae after vertical and horizontal infection with the newly characterized tilapia larvae encephalitis virus (TLEV) was accessed by evaluation of cell-mediated reactive oxygen species (ROS) production in affected fish with the use of horseradish peroxidase-amplified luminol-dependent chemiluminescence assay. The priming in-vivo infection with TLEV resulted in downregulation of ROS response in both vertically- and horizontally-infected fish; this suppression was further exacerbated by specific in-vitro booster infection with the same virus. Application of Ca ionophore and phorbol myristate acetate as alternative nonspecific boosters enabled restoration of ROS release in vertically-infected but not in horizontally-infected larvae.

Viral encephalitis of tilapia larvae: primary characterization of a novel herpes-like virus.

We report here an outbreak of an acute disease that caused high mortality rate in laboratory-reared tilapia larvae. The disease was initially observed in inbred gynogenetic line of blue tilapia larvae (Oreochromis aureus) and could be transmitted to larvae of other tilapia species. Based on the clinical manifestation (a whirling syndrome), we refer to the disease as viral encephalitis of tilapia larvae.

Determinants of pathology in light-irradiated cells.

This work is devoted to further evaluation of pathological states in light-irradiated cells in terms of relationship between the level of reactive oxygen species (ROS) and polymorphonuclear neutrophil (PMN) count. Human leukocytes were isolated from 34 donors exhibiting flu-like symptoms, irradiated with a nonlaser blue light (2 and 5 mW cm(-2)) and examined for ROS production by a chemiluminescence (CL) assay. The level of ROS was measured as a basal chemiluminescence before (BCL0) and after (BCL) irradiation.

Male-specific protein (MSP): a new gene linked to sexual behavior and aggressiveness of tilapia males.

MSP is a male-specific protein initially identified in the serum of sexually active Sarotherodon galilaeus males, and is shown herein to be present in the serum of sexually mature males, but not females, of three other tilapia species. Cloning of the MSP cDNA and analysis of its predicted amino-acid sequence revealed that it is an outlier lipocalin that contains a signal peptide in its N-terminal region. The abundance of highly homologous sequences found in fish and the monophyletic relationship to tetrapod Alpha-1-acid glycoprotein (AGP) places it as a clade XII lipocalin.

Immunogenicity of bioactive magnetic nanoparticles: natural and acquired antibodies.

Uniform magnetic nanoparticles (MNP) were prepared by nucleation followed by controlled growth of maghemite thin films onto porcine gelatin nuclei. The formed gelatin containing MNP (Gel-MNP) were then coated with dextran (Gel-MNP-Dex) followed by human serum albumin (Gel-MNP-Dex-HSA). Since these MNP are designated for clinical applications, studies concerning the immunogenicity of their antigenic components (porcine gelatin, dextran, and HSA) have been performed in BALB-C mice.

Chemiluminescent analysis of light-irradiated leukocytes as a diagnostic tool for fast identification of pathological states.

Objective: Reactive oxygen species (ROS), mainly produced by polymorphonuclear neutrophils (PMN), are a significant part of host defense in pathologic states. We attempted to relate numbers of PMN and ROS generated within PMN to develop an alternative photochemical approach for evaluation of the potential of these cells to resist the development of inflammatory pathology.

Background Data: Lack of sensitivity to light has been reported in healthy cells, while sensitivity to light characterizes cell pathology.

Nano- and microparticles as adjuvants in vaccine design: success and failure is related to host natural antibodies.

Bovine serum albumin (BSA) and the surface A-layer protein (AP) of an atypical strain of fish bacterial pathogen Aeromonas salmonicida were covalently linked with polymeric nano- and microparticles, and antigenicity of the resulted conjugates was compared in mice and goldfish. Distinct albeit different levels of natural BSA and AP antibodies were present in both animal species. Significant stimulation of the anti-AP antibody response in mice strikingly contrasted to unresponsiveness or even suppression in fish.

Altered self-erythrocyte recognition and destruction in an inbred line of tilapia (Oreochromis aureus).

Carboxyfluorescein diacetate (cFDA)-stained autologous and syngeneic tilapia (Oreochromis aureus) erythrocytes are recognized by effector peripheral blood leukocytes and lysed after a short culture period of 4 h. The hemolysis level was evaluated by measuring the fluorescence of the released cFDA. The degree of lysis of stained target erythrocytes of 60 individuals revealed a trimodal distribution statistically stratified into three groups of low (LR), intermediate (IR), and high (HR) responders.

Influence of visible light and ultraviolet irradiation on motility and fertility of mammalian and fish sperm.

Objective: The effects of visible light irradiation on sperm motility, fertility, and reactive oxygen species (ROS) formation were investigated and compared in ram and fish (tilapia).

Background Data: Low-energy visible light has previously been found to modulate various processes in different biological systems. In the literature, it is accepted that the first step following visible light irradiation is the formation of ROS by endogenous cellular photosensitizers.

Experimental handling stress as infection-facilitating factor for the goldfish ulcerative disease.

Experimental handling stress (EHS) was applied to clinically asymptomatic farmed goldfish (Carassius auratus L.). EHS affected the gills and skin integrity of the fish and was accompanied by increased levels of plasma glucose, cortisol and interleukin-10 (IL-10).

Upregulation of carp GDNF mRNA by the immunomodulator AS101.

Although the glia derived neurotrophic factor (GDNF) is defined as a molecule that maintains neuronal cells, it possesses a range of functions outside the nervous system. For example, it is essential for uretric branching in kidney morphogenesis and for regulating the differentiation of stem cells during spermatogenesis, cardiac, hair follicle and vascular differentiation and the maintenance of immune cells. In the present work, the presence of GDNF in carp peripheral blood leukocytes (PBL) and head kidney cells (HK) was evidenced and its evolutionary importance in both neural and immune systems development was suggested.

Immunisation of fish by bath immersion using ultrasound.

Ultrasonic irradiations (USI) as a means to open routes in the skin, thus facilitating the transdermal delivery of vaccines that will improve the effectiveness of vaccination by immersion, are reviewed in this paper. Based on our recent results in goldfish and carp it could be summarized that: (i) USI significantly improved the antigen uptake and enhanced antibody response; (ii) the requirements for high antigen concentrations, which are needed for simple bath immersion, could be considerably reduced in presonicated fish; (iii) after bath immersion, the antigen was slowly released from the skin to the blood in which its presence could still be detected 24 hours later. This retardation of the antigen in the skin was suggested to be due to a possible interaction with cells of the local immune system, in which it is processed and recognized.

Visible light affects chemiluminescence of carp (Cyprinus carpio) blood leukocytes.

Objective: Visible light irradiations at doses of 5 and 12 J/cm(2) were applied to carp buffy coat leukocytes.

Materials And Methods: The leukocytes response was measured by a chemiluminescence (CL) assay as basal (spontaneous) bCL and Ca ionophore-induced stimulated CL (StCL).

Results: The irradiation caused a significant decrease in bCL in six out of 14 fish (susceptible fish) and rendered eight out of 14 fish unsusceptible.

Enhancement of antigen uptake and antibody production in goldfish (Carassius auratus) following bath immunization and ultrasound treatment.

Ultrasound irradiation and hyperosmotic treatments were compared as facilitators of antigen (BSA) penetration through the skin by bath vaccination and as enhancers of the antibody response in goldfish. The kinetics of BSA penetration and accumulation into the skin, and via it to the blood, and the consequent specific stimulation of the humoral immune response, were studied. The main findings are: (1).

Monitoring the apoptotic process induced by oxidized low-density lipoprotein in Jurkat T-lymphoblast and U937 monocytic human cell lines.

Cell death is a major event in the pathophysiology of atherosclerosis. Oxidized low-density lipoprotein (Ox-LDL), which plays a key role in the atherogenesis, has a powerful cytotoxic effect and causes necrosis or apoptosis of different types of cells. In the present work we studied the mechanism of cell death in two model systems: T lymphocytes and monocytes cell line, exposed to Ox-LDL.

betaFSH, betaLH and growth hormone gene expression in blue gourami (Trichogaster trichopterus, Pallas 1770) during spermatogenesis and male sexual behavior.

The relationship between gonadal development (histological evidence for spermiogenesis and/or spermatogenesis), sexual behavior (nest-building) and mRNA levels of gonadotropins (betaFSH and betaLH) and growth hormone (GH) in the male pituitary was investigated. Amplification of betaFSH cDNA showed a significantly higher mRNA level in mature males (whether sexually active or not) than in juveniles. However, following PCR amplification of betaLH cDNA, a significantly higher mRNA level was found in the sexually active group compared to the sexually inactive group.

Association between loci with deleterious alleles and distorted sex ratios in an inbred line of tilapia (Oreochromis aureus).

Three microsatellite markers (UNH159, UNH231, and UNH216) were examined for association with both deleterious genes and sex-ratio distortions in a full-sib family of 222 progeny from the fourth generation of a meiogynogenetic tilapia line (Oreochromis aureus). The three markers were mapped previously to different linkage groups and were shown to be associated with genes with deleterious alleles in this line. A restricted maximum likelihood model was used for analysis of major effects and their interactions on sex ratio and viability.

Natural antibodies and their significance in active immunization and protection against a defined pathogen in fish.

Natural antibody activity against Aeromonas salmonicida extracellular A-layer protein (A-protein) showed large individual variations in a farmed group of 101 goldfish (Carassius auratus L.). Statistical analyses of these variations led us to divide this group into homogeneous high and low naturally active (HNA and LNA) subgroups.