Cyprinid herpesvirus-3 (CyHV-3) disturbs osmotic balance in carp (Cyprinus carpio L.)--A potential cause of mortality.

Cyprinid herpesvirus-3 (CyHV-3) causes a fatal disease in carp (Cyprinus carpio) and its ornamental koi varieties which seriously affects production and trade of this fish species globally. Up to now, the pathophysiology of this disease remains unclear. Affected individuals develop most prominent lesions in gills, skin and kidney, in tissues which are involved in the osmotic regulation of freshwater teleosts. Therefore, here serum and urine electrolyte levels were examined during the course of an experimental infection of carp with CyHV-3. In infected carp an interstitial nephritis with a progressive deterioration of nephric tubules developed, which was paralleled by elevated electrolyte losses, mainly Na(+) in the urine. The urine/plasma ratio for Na(+) increased from 0.03 in uninfected carp to 0.43-0.83 in carp under CyHV-3 infection, while concentration of divalent ions were not significantly changed. These electrolyte losses could not be compensated since plasma osmolality and Na(+) concentration dropped significantly in CyHV-3 infected carp. This was most probably caused by the progressive deterioration of the branchial epithelium, which in teleosts plays a prominent role in osmoregulation, and which was seen concomitantly with decreasing electrolyte levels in the serum of carp under CyHV-3 infection. Immediately after infection with CyHV-3, by day 2 post exposure, affected carp showed severe anaemia and prominent leucocytosis indicating the development of an acute inflammation, which could intensify the observed hydro-mineral imbalances. The data presented here show that an infection with CyHV-3 induces an acute inflammation and a severe dysfunction of osmoregulation in affected carp or koi, which may lead to death in particular in the case of acute disease progression.

Changes in skin mucus of common carp, Cyprinus carpio L., after exposure to water with a high bacterial load.

Water in aquaculture systems may contain a high load of microorganisms. Reduction in overall bacterial tank water load improves fish health and growth parameters. In this study, the effect of an increase of overall bacterial load in tank water on carp skin mucus was assessed. Intracellular and released high molecular weight glycoproteins (HMGs) of carp skin mucus were analysed for changes using histological, histochemical and biochemical techniques. Increase of bacterial load did not induce obvious clinical responses in carp, but the skin of exposed carp responded quickly. The amount of skin mucus HMGs isolated increased as well as their total glycosylation. An increased goblet cell number was observed for all carbohydrate stainings, but most clearly for acidic glycoconjugates. A change in the terminal presence of some sugars was also seen. After the initial response of carp, an adaptation to the higher bacterial load in the water appeared to occur as mucins had a higher glycosylation. The changes observed suggest that these skin mucus adaptations are part of a primary defence mechanism of mucosal epithelia, even at a low pathogenic pressure.

Influence of carp intestinal mucus molecular size and glycosylation on bacterial adhesion.

The first step of the pathogenesis of many infectious diseases is the colonisation of the mucosal surface by the pathogen. Bacterial colonisation of the mucosal surface is promoted by adherence to high molecular weight mucus glycoproteins. We examined the effect of carp intestinal mucus glycoproteins on the adhesion of different bacteria. The bacteria used were 3 strains of Aeromonas hydrophila, and A. salmonicida, Edwardsiella tarda and Yersinia ruckeri. All bacteria adhered to mucus, but at varying intensities. All tested bacteria adhered best to molecules of 670 to 2000 kDa in size, less to molecules larger than 2000 kDa and weakest to molecules of 30 to 670 kDa. In general, bacteria that showed a stronger adhesion to intestinal mucus were cytotoxic to cells in vitro, and bacteria that showed a weaker adhesion to intestinal mucus did not lead to alterations of monolayers of EPC-cells. Furthermore, the involvement of glycan side chains of the glycoproteins for bacterial adhesion was analysed for one A. hydrophila strain. After cleavage of terminal sugar residues by treatment of mucus glycoproteins with different glycosidases, binding of bacteria was modulated. When mannose was cleaved off, adhesion significantly increased. Blocking of glycan receptors by incubation of bacteria with different oligosaccharides had no clear effect on bacterial binding to mucus glycoproteins. Our results suggest that bacteria interact with carbohydrate side chains of mucus glycoproteins, and that the carbohydrates of the core region are involved in bacterial binding.

Chemotaxis towards, adhesion to, and growth in carp gut mucus of two Aeromonas hydrophila strains with different pathogenicity for common carp, Cyprinus carpio L.

Characteristics that promote bacterial colonization of the intestinal mucosal surface were examined in two strains of the common fish pathogen Aeromonas hydrophila, with different pathogenicity. The characteristics examined were chemotactic activity towards mucus, bacterial adherence to mucus and growth in mucus. Intestinal gut mucus of healthy common carp was used. The results indicate that chemotaxis is not necessary for a bacterium to become pathogenic, but it may be a necessary parameter for a bacterium to be an obligate pathogen. Adhesion also seems to be a factor influencing pathogenicity. The results suggest that higher adhesion to mucus and subsequent growth is associated with differences in pathogenicity.