Hepatocellular adaptation to extreme nutritional conditions in ide,Leuciscus idus melanotus L. (Cyprinidae). A morphofunctional analysis

The adaptive response of the omnivorous ide,Leuciscus idus melanotus, to drastic metabolic conditions was analyzed on different levels of organisation investigating a variety of parameters: Organism (condition factor, liver-somatic index), organ (liver structure), cellular and subcellular level (hepatocyte structure, glycogen and lipid storage, contents and distribution of nucleic acids, enzyme alterations).During starvation, ide were able to maintain liver integrity in a biphasic process: after an initial phase of disturbance, ide established a new structural and metabolic homeostasis. Recovery from starvation was possible only with a complete diet but not with a sucrose diet. Carbohydrate overload, as evoked by sucrose refeeding, did not result in liver or carcass fattening as known from mammals.To the best of our knowledge, the present study is the first to use enzyme histochemistry in fish nutrition research. In mammals, histochemistry is particularly helpful for understanding processes of hepatic metabolic adaptation. In fish, however, on the basis of our results, enzyme histochemical studies appear to be of limited value, as long as no further data are available on a zonal distribution of enzyme activities in teleost liver parenchyma. Instead, the histochemical detection of the distribution of hepatic storage products and RNA-positive material yielded important information on liver adaptive processes.Abbreviations H starvation - SU sucrose - AD artificial diet - AcP acid phosphatase - G6Pase glucose-6-phosphatase - G6PDH glucose-6-phosphate dehydrogenase - GLU -glucuronidase - ME malic enzyme - PHO glycogen phosphorylase - UE unspecific esterase - RER rough endoplasmic reticulum     

Binding and bioactivity of insulin in primary cultures of carp (Cyprinus carpio) hepatocytes

Isolated carp hepatocytes cultured in serum-free, chemically defined medium were used to investigate within the same cell preparation characteristics of the binding of insulin as well as effects of insulin on cellular metabolism. The binding of human [¹²⁵I]-insulin to carp hepatocytes was studied in kinetic, saturation and displacement experiments. A dependency of insulin binding on the collagenase used for cell isolation was demonstrated. Insulin binding decreased during the first 12h of culture but remained constant during the following 12h. The kinetic experiments revealed that [¹²⁵I]-insulin binding reached a steady state within 20–30 min of incubation. The mathematical analysis of the saturation experiments demonstrated the existence of two populations of binding sites, one with high affinity (K_d1 = 5.5 pM) and low capacity (B_max1 = 0.14 fmol/mg protein or 77 binding sites/cell) and one with low affinity (K_d2 = 2.4 nM) and high capacity (B_max2 = 17.6 fmol/mg protein or 9623 binding sites/cell). In competition experiments, 312 pM [¹²⁵I]-insulin was displaced by cold insulin, IGF-I and IGF-II with IC₅₀ values of 2.2, 7.9 and 20.3 nM, respectively. Glucagon was without effect. Binding of insulin to carp hepatocytes resulted in a significant reduction of glucose release and a significant increase of protein synthesis as of de novo fatty acid synthesis. dedicated to Prof. Dr. W. Hanke on the occasion of his 65^th birthday.     

Investigations into the temporal development of epitheliocystis infections in brown trout: a histological study

Epitheliocystis in Swiss brown trout (Salmo trutta) is a chlamydial infection, mainly caused by Candidatus Piscichlamydia salmonis and Candidatus Clavichlamydia salmonicola. To gain a better understanding of the temporal development of infections in wild brown trout, we investigated epitheliocystis infections during the course of the summer and autumn months of a single year (2015), and compared this to sampling points over the span of the years 2012–2014. The survey focused on tributaries (Venoge and Boiron) of the Rhone flowing in to Lake Geneva. When evaluated histologically, epitheliocystis infections were found throughout the period of investigation with the exception of the month of June. Fifty to 86 animals per sampling were investigated. Highest prevalence and infection intensities were seen in September. A correlation between epitheliocystis infection and water temperatures was not evident. Interyear comparison revealed consistent levels of prevalence and infection intensities in late summer. The absence of infections in June, combined with the consistent interyear results, indicates seasonal fluctuation of epitheliocystis infections in brown trout with a reservoir persisting during winter months from which infections can re‐initiate each year. This could either be at levels below detection limits within the brown trout population itself or in an alternative host.     

Are brown trout Salmo trutta fario and rainbow trout Oncorhynchus mykiss two of a kind? A comparative study of salmonids to temperature‐influenced Tetracapsuloides bryosalmonae infection

Proliferative kidney disease (PKD) of salmonids caused by Tetracapsuloides bryosalmonae causes high mortalities of wild brown trout (Salmo trutta fario) and farmed rainbow trout (Oncorhynchus mykiss) at elevated water temperatures. Here the aim was to compare the temperature‐dependent modulation of T. bryosalmonae in the two salmonid host species, which display different temperature optima. We used a novel experimental set‐up in which we exposed brown trout and rainbow trout to an identical quantified low concentration of T. bryosalmonae for a short time period (1 hr). We followed the development of the parasite in the fish hosts for 70 days. PKD prevalence and parasite kinetics were assessed using qPCR. Exposures were performed at temperatures (12°C and 15°C) that reflect an environmental scenario that may occur in the natural habitat of salmonids. T. bryosalmonae infection was confirmed earliest in brown trout kept at 15°C (day 7 post‐exposure) while, in all other groups, T. bryosalmonae was not confirmed until day 15 post‐exposure. Moreover, significantly greater infection prevalence and a faster increase of parasite intensity were observed in brown trout kept at 15°C than in all other groups. These results indicate that PKD is differentially modulated by water temperature in related host species.     

Health of farmed fish: its relation to fish welfare and its utility as welfare indicator

This brief review focuses on health and biological function as cornerstones of fish welfare. From the function-based point of view, good welfare is reflected in the ability of the animal to cope with infectious and non-infectious stressors, thereby maintaining homeostasis and good health, whereas stressful husbandry conditions and protracted suffering will lead to the loss of the coping ability and, thus, to impaired health. In the first part of the review, the physiological processes through which stressful husbandry conditions modulate health of farmed fish are examined. If fish are subjected to unfavourable husbandry conditions, the resulting disruption of internal homeostasis necessitates energy-demanding physiological adjustments (allostasis/acclimation). The ensuing energy drain leads to trade-offs with other energy-demanding processes such as the functioning of the primary epithelial barriers (gut, skin, gills) and the immune system. Understanding of the relation between husbandry conditions, allostatic responses and fish health provides the basis for the second theme developed in this review, the potential use of biological function and health parameters as operational welfare indicators (OWIs). Advantages of function- and health-related parameters are that they are relatively straightforward to recognize and to measure and are routinely monitored in most aquaculture units, thereby providing feasible tools to assess fish welfare under practical farming conditions. As the efforts to improve fish welfare and environmental sustainability lead to increasingly diverse solutions, in particular integrated production, it is imperative that we have objective OWIs to compare with other production forms, such as high-density aquaculture. However, to receive the necessary acceptance for legislation, more robust scientific backing of the health- and function-related OWIs is urgently needed.     

PKD: the proliferative kidney disease in fish

The present study gives an overview about the knowledge and the diagnostic possibilities of the proliferative kidney disease (PKD) in fishes. This disease is responsible for economically significant losses in farmed fish and severe reductions in wild fish populations in Europe and North America. PKD is affecting mainly salmonid species. In Swiss rivers the proliferative kidney disease pictures the highest percentage of diseases among brown trouts. The main symptom of PKD is a massive proliferation of the interstitial kidney tissue which named the disease. Other unspecific symptoms are anemia, ascites, exophthalmus and apathy. The causative agent of the disease is Tetracapsula bryosalmonae (Myxozoa). Many aspects of the life cycle of this parasite remain unclear so far but it is known that the appearance of this parasite is linked to the presence of a range of freshwater bryozoans. PKD appears mostly in summer during high water temperatures (> or = 15 degrees C) and can lead to high mortality rates. A diagnosis can be made by using light-microscopic, lectinimmunhistochemical as well as molecular biological methods. Before we can develop successful management strategies the knowledge gaps in understanding the disease cycle and the exact pathogenesis of PKD need to be closed.     

Post-prandial protease activity in the digestive tract of African catfish Clarias gariepinus larvae fed decapsulated cysts of Artemia

The alkaline proteolytic activity in the gut of African catfish larvae was studied during short time ranges from 30 min to 4 h after ingestion of decapsulated Artemia cysts. The variation in total protease and trypsin activities during the day was monitored during starvation, after one single meal ingestion, and during continuous feeding. In starved larvae the enzymatic activity was low and did not change in time. No significant endogenous secretion of digestive enzymes was detected. The level of alkaline proteolytic activity found in starved larvae was further considered as the basal level. In larvae fed only one meal during the day, the enzyme activity significantly increased from 3 h post-feeding up to a maximum level found 12 h after feeding. In the larvae receiving a meal every 4 h, the effect of feeding on the proteolytic activity was significantly different from the one in fish fed only once a day. The total protease activity in this dietary treatment changed according to the time of feeding and fluctuated around a constant level, which was intermediate between the maximum and the basal level. No rhythmic cycle of enzyme production in the fish was observed when the proteolytic activity was studied during a cycle of 24 h. When specific trypsin activity was measured, a similar pattern was found as with the total protease. The contribution of digestive enzymes from Artemia to the total digestion of food by the catfish larvae was calculated to be less than 1% of the total amount of the proteolytic activity measured in the larval gut.     

Heat treatment affects protein quality and protease activity in decapsulated cysts of Artemia when used as starter food for larvae of African catfish Clarias gariepinus (Burchell)

Decapsulated cysts of Artemia subjected to different heat treatments (40, 60, 80 and 96 °C) were fed to African catfish Clarias gariepinus larvae. Heated cysts, untreated cysts and live Artemia nauplii as control constituted the experimental diets. Protein denaturation and solubility, total alkaline protease and specific trypsin activities in the cyst diets were evaluated. The growth of catfish larvae and the proteolytic activity of larval samples during development were also determined. Heat treatment of cysts increased protein denaturation and decreased protein solubility. The protease activity in the cyst diets decreased with higher heating temperatures. The growth of catfish larvae differed according to the diet; higher fish growth was achieved with nauplii and cysts heated at 40 °C. The digestive enzyme activity in larval samples remained similar in all dietary treatments during larval development. The quality of food protein and the way this protein is processed might be more important for successful larval growth than exogenous enzyme supply.     

Proliferative kidney disease in Switzerland: current state of knowledge

This study presents an overview of the distribution of proliferative kidney disease (PKD) in wild and farmed fish in Switzerland based on two sources: (1) the results of routine diagnostic work at the National Fish Disease Laboratory, and (2) the results of a country‐wide survey for PKD. The first case of PKD in Switzerland was diagnosed in 1979 and since 1981, a few cases have been found every year. Affected species were rainbow trout, brown trout and grayling from rivers and fish farms. Most of the diseased fish were found in the lower altitude regions of the country (Swiss midlands). A seasonal distribution of the cases was evident: PKD‐positive fish occurred only from June to November withprominent peaks in August and September. Among infected fish all length classes were present, but the highest numbers of affected fish were found in the length classes <10 cm and 16–20 cm. The evaluation of the samples from a survey of the occurrence of PKD in 2000/2001 revealed a much broader geographical distribution of the infection than expected from routine diagnostic observations. Fish from 56 of 139 rivers analysed by histological examination were positive for parasites. Fish from 131 of these sites were examined macroscopically. Of these, trout from 45 sites showed gross alterations indicative for PKD. Most of the positive fish were found in the Swiss midlands north of the Alps. Fish from six of 65 fish farms investigated were found to be PKD‐positive by histopathological analysis, and in four of these cases fish could be identified as infected on the basis of gross examination.