■ Abstract Apoptosis is a genetically programmed, morphologically distinct form of cell death that can be triggered by a variety of physiological and pathological stimuli. Studies performed over the past 10 years have demonstrated that proteases play critical roles in initiation and execution of this process. The caspases, a family of cysteine-dependent aspartate-directed proteases, are prominent among the death proteases. Caspases are synthesized as relatively inactive zymogens that become activated by scaffold-mediated transactivation or by cleavage via upstream proteases in an intracellular cascade. Regulation of caspase activation and activity occurs at several different levels: ( a) Zymogen gene transcription is regulated; ( b) antiapoptotic members of the Bcl-2 family and other cellular polypeptides block proximity-induced activation of certain procaspases; and ( c) certain cellular inhibitor of apoptosis proteins (cIAPs) can bind to and inhibit active caspases. Once activated, caspases cleave a variety of intracellular polypeptides, including major structural elements of the cytoplasm and nucleus, components of the DNA repair machinery, and a number of protein kinases. Collectively, these scissions disrupt survival pathways and disassemble important architectural components of the cell, contributing to the stereotypic morphological and biochemical changes that characterize apoptotic cell death.

Mammalian caspases: structure ,a ctivation ,s ubstrates, and functions during apoptosis

Innate immunity of fish (overview).

Fish welfare: Current issues in aquaculture

A review of the main bacterial fish diseases in mariculture systems.

All metazoans possess innate immune defence system whereas parameters of the adaptive immune system make their first appearance in the gnathostomata, the jawed vertebrates. Fish are therefore the first animal phyla to possess both an innate and adaptive immune system making them very interesting as regards developmental studies of the immune system. The massive increase in aquaculture in recent decades has also put greater emphasis on studies of the fish immune system and defence against diseases commonly associated with intensive fish rearing. Some of the main components of the innate and adaptive immune system of fish are described. The innate parameters are at the forefront of immune defence in fish and are a crucial factor in disease resistance. The adaptive response of fish is commonly delayed but is essential for lasting immunity and a key factor in successful vaccination. Some of the inherent and external factors that can manipulate the immune system of fish are discussed, the main fish diseases are listed and the pathogenicity and host defence discussed. The main prophylactic measures are covered, including vaccination, probiotics and immunostimulation. A key element in the immunological control of fish diseases is the great variation in disease susceptibility and immune defence of different fish species, a reflection of the extended time the present day teleosts have been separated in evolution. Future research will probably make use of molecular and proteomic tools both to study important elements in immune defence and prophylactic measures and to assist with breeding programmes for disease resistance.

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Immunological Control of Fish Diseases

Prevention of stress in rainbow trout (Oncorhynchus mykiss) fed diets containing different doses of glucan

Over the years, aquaculture has shown increasing development in terms of production. However, due to intensive farming practices, infectious diseases represent the main problem in fish farms, causing heavy economic losses. The use of antibiotics for controlling diseases is widely criticized for its negative impact, including selection of antibiotic-resistant bacterial strains, immunosuppression, environmental pollution and accumulation of chemical residues in fish tissues. On the other hand, though vaccination is the most effective prophylactic method of preventing disease outbreaks, the development of effective formulations is often hindered by high production costs and the antigenic heterogeneity of the microbial strains. Recently, there has been increased interest in the possibility of using medicinal herbs as immunostimulants, capable of enhancing immune responses and disease resistance of cultured fish. Plant-derived products seem to represent a promising source of bioactive molecules, being at the same time readily available, inexpensive and biocompatible. The aim of this article is to provide an overview of recent research dealing with the use of medicinal plants in aquaculture. Special attention is given to the information about the effects of plant extracts/products on fish growth, haematological profiles, immune responses and resistance to infectious diseases.

Current research on the use of plant-derived products in farmed fish

The research was aimed to assess the effect of dietary carvacrol (0.025% and 0.05%) on sea bass (Dicentrarchus labrax) growth, immune response and resistance to Listonella anguillarum. Fish (69.2 ± 0.22 g) were fed the experimental diets for 9 weeks. Dietary carvacrol did not negatively affect fish survival, growth performance, feed intake and feed conversion ratio (P > 0.05) nor carcass yield and viscerosomatic, hepatosomatic and mesenteric fat index (P > 0.05). Serum and head kidney leucocytes were collected after 1, 4 and 8 weeks of feeding. Carvacrol significantly reduced serum proteins, immunoglobulins and lysozyme activity (P < 0.01) and moderately increased phagocytosis and pinocytosis of head kidney macrophages. The release of reactive oxygen species by leucocytes was reduced in carvacrol-fed fish, even if significantly (P < 0.05) only in those fed 0.05% carvacrol for 1 week. Dietary carvacrol did not significantly affect the aspecific immune response, although a potential antioxidant activity might be speculated. Moreover, feeding carvacrol provided an appreciable resistance to a challenge with L. anguillarum, when a bacterial dose lower than the Lethal Dose50 was used. Cumulative mortality in fish fed 0.025% carvacrol was significantly lower than that of untreated controls (75% Relative Per cent Survival).

Growth parameters, innate immune response and resistance to Listonella (Vibrio) anguillarum of Dicentrarchus labrax fed carvacrol supplemented diets

Bacterial cells of the marine fish pathogen Photobacterium damsela subsp. piscicida were grown in novel culture media. A mixture of whole cells and extracellular components was inactivated and used in bath, intraperitoneal (i.p.) and oral vaccination of sea bass, Dicentrarchus labrax, employing two sizes of fish. A commercial vaccine was used for comparative purposes. Control and immunized fish were either bath or intraperitoneally challenged 6 and 12 weeks post-vaccination. Small fish had significantly higher relative percentage survival with the novel vaccine mixture both at 6 and 12 weeks post-vaccination by bath, in comparison with the commercial vaccine. No protection was afforded at 6 or 12 weeks post-immunization by either vaccine after challenge via i.p. injection. Sea bass (1.5-2 g) intraperitoneally vaccinated with various adjuvanted vaccine mixtures were not protected against pasteurellosis. In contrast, larger sea bass (20 g) benefited from vaccination with the novel vaccine mixtures. Intraperitoneal challenge with the pathogen resulted in protection in both fish groups vaccinated with novel vaccine mixtures, whereas control fish suffered high mortalities (> 80%). Orally vaccinated fish were immersion challenged with the pathogen. At 6 and 12 weeks post-vaccination the control fish had a high mortality and the fish vaccinated with the novel vaccine mixture achieved good protection.

Vaccination trials of sea bass, Dicentrarchus labrax (L.), against Photobacterium damsela subsp. piscicida, using novel vaccine mixtures.

Study design Dorsal arthrodesis of thoracic spine in a prepubertal New Zealand White rabbit model Objective Evaluating the consequences of dorsal arthrodesis on the growth of the spine, sternum, and thorax in prepubertal rabbits, through the study of CT scans Summary of background data Vertebral arthrodesis in the treatment of progressive idiopathic scoliosis in prepubertal patients is not ideal, but is still a choice in treating major deformities of the spine Postoperative assessment of spinal deformity is essential, feasible, and recordable through CT scans Methods Twelve female rabbits, 9 weeks old, were subjected to surgery for dorsal arthrodesis of the upper thoracic spine Surgery involved the implant of 2 "C"-shaped titanium bars, which were placed beside the spinous processes of the thoracic vertebrae Three CT scans were performed, 10 (T1), 55 (T2), and 139 (T3) days after surgery Measures were obtained by Myrian Pro software for 3 different groups: G1 with complete fusion, G2 with incomplete fusion, and G3 sham-operated Results The average of the dorsoventral/laterolateral thoracic diameter ratio at fused levels is lower than 1 in G1 as well as in G2; on the contrary, in G3 is higher than 1 The average growth of the sternum length between T1 and T2 and between T2 and T3 is minor in G1 than in G2 and G3 The dorsal and ventral lengths of thoracic vertebral bodies in the spinal segment D1-D6 is smaller in G1 and G2 than in G3, whereas no differences were observed between the 3 groups in the D7-D12 segment without arthrodesis Conclusion Dorsal arthrodesis in prepubertal rabbits changes thoracic growth patterns In operated rabbits, the dorsoventral thoracic diameter grows more slowly than the laterolateral thoracic diameter The sternum as well as the lengths of thoracic vertebral bodies in the spinal segment D1-D6 grow less The crankshaft phenomenon is evident at the fused vertebral levels where there is a reduction of thoracic kyphosis

Donatella Volpatti

Papers

Prevention of stress in rainbow trout (Oncorhynchus mykiss) fed diets containing different doses of glucan

Current research on the use of plant-derived products in farmed fish

Growth parameters, innate immune response and resistance to Listonella (Vibrio) anguillarum of Dicentrarchus labrax fed carvacrol supplemented diets

Vaccination trials of sea bass, Dicentrarchus labrax (L.), against Photobacterium damsela subsp. piscicida, using novel vaccine mixtures.

Dorsal arthrodesis of thoracic spine and effects on thorax growth in prepubertal New Zealand white rabbits.