Ecological influences on the difference in δ15N and δ13C values between fish tissues: implications for studies of temporal diet variation

Temporal diet changes can be detected by comparing δ¹³C and δ¹⁵N values between tissues with different isotopic turnover times. However, other factors contribute to disparities in δ¹³C and δ¹⁵N signatures between tissues and could confound the interpretation of stable isotope data. We examined the effects of ecological factors on differences in muscle and liver δ¹³C and δ¹⁵N signatures of Arctic charr [Salvelinus alpinus, (L.)] to determine whether their effects were large enough to influence studies of diet change. In addition, we examined whether differences in lipid content accounted for a significant portion of the differences in δ¹³C values between tissues. Results indicated that life history, diet, reproductive status and gender had significant effects on the differences in δ¹³C values between tissues, while gender and diet significantly affected differences in δ¹⁵N values. Differences in % lipid content between tissues also explained approximately 74% of the observed variation in the difference in δ¹³C values between tissues. The differences in δ¹³C values observed between muscle and liver tissues may be partially explained by physiological differences in lipid storage and use between tissues associated with ecological factor effects. These results demonstrate that ecological factors other than a change in diet can influence the differences in δ¹³C and δ¹⁵N signatures between muscle and liver tissues. However, the impact of ecological factors on studies of diet change depends on how the magnitude of the factor effects compared with a biologically meaningful change in diet.     

Effect of Modified Atmosphere Packaging on Quality and Shelf Life of Salted Bonito (Sarda sarda)

In this study, the fresh meat of bonito (Sarda sarda) was used to make lakerda, which is a type of salted fish (Turkish traditional product). The lakerda was packaged in plastic film (A), vacuumed (VP), placed in modified atmosphere packaging (MAP; M1 = 80%/20% CO₂/N₂, M2 = 65%/35% CO₂/N₂, M3 = 30%/60%/10% CO₂/N₂/O₂), and stored at 2 ± 1°C. Microbiological, chemical, sensory, and color analyses were conducted to determine the changes in quality. Total viable bacteria counts did not exceed the limit of consumption (7 log cfu/g) for all groups on Day 31. Total volatile basic nitrogen (TVB-N) and trimethylamine- nitrogen (TMA-N) values of M1, M2, and M3 remained lower than the proposed acceptability limits of 35 and 12 mg N/100 g, respectively, up to 31 days of storage. However, VP lakerda samples exceeded TVB-N limits after 31 days of storage, whereas A lakerda samples exceeded after 8 days of storage. In our study, sensory analysis determined the shelf life of modified atmosphere packaged lakerda. When Group A is compared with the other groups, MAP and VP lengthened the shelf life of lakerda by 23 days.     

The use of stable isotope ratios δ13C and δ15N to track the incorporation of Ulva and other important dietary ingredients into the gonads of the sea urchin Tripneustes gratilla

In this study, we demonstrate how stable carbon (C) and nitrogen (N) isotope analysis can be used to estimate the allocation of specific dietary ingredients, incorporated into four artificially formulated feeds supplemented with varying amounts of dried seaweed Ulva armoricana (0, 50, 150 and 200 g/kg), to the production of gonads by Tripneustes gratilla over a 20‐week growth trial. Results indicate that U. armoricana is an important dietary ingredient for gonad production, accounting for an average of 37.33 ± 1.03% of the isotopic signal recorded for gonads at the end of the growth trial. This is significantly more than fish meal and soya, which only contributed an average of 11.40 ± 1.81 and 3.80 ± 0.09%, respectively, of the isotopic signal of the gonads. Maize and wheat each account for almost 25% of all the dietary ingredients used to formulate the feeds, but their relative contribution to gonad production only averaged 8.55 ± 0.58 and 3.80 ± 0.05%, respectively, among the diets. This indicates that, at least for gonad development, these ingredients may not be required at such high levels. These findings support previous data on the importance of Ulva in abalone and urchin diets, and demonstrate the value of stable isotope analysis for assessing the contribution of specific dietary ingredients in new feed formulations, particularly where growth of specific tissues is being investigated.     

Effect of starvation and refeeding on the hepatopancreas of whiteleg shrimp Penaeus vannamei (Boone) using computer‐assisted image analysis

Under normal farming conditions, shrimp can experience starvation periods attributable to disease outbreaks or adverse environmental conditions. Starvation leads to significant morphological changes in the hepatopancreas (HP), being the main organ for absorption and storage of nutrients. In the literature, limited research has described the effect on the HP of periods of starvation followed by refeeding and none in whiteleg shrimp (Penaeus vannamei) using computer‐assisted image analysis (CAIA). This study describes the effect of starvation and starvation followed by refeeding on the HP of whiteleg shrimp using CAIA. Visiopharm^® software was used to quantify the following morphological parameters, measured as ratio to the total tissue area (TLA): total lumen area (TLA:TTA), haemocytic infiltration area in the intertubular spaces (HIA:TTA), B‐cell vacuole area (VBA:TTA), lipid droplet area within R cells (LDA:TTA) and F‐cell area (FCA:TTA). Significant changes were measured for HIA:TTA and LDA:TTA during starvation (increase in HIA:TTA associated with decrease in LDA:TTA) and starvation followed by refeeding (decrease in HIA:TTA associated with increase in LDA:TTA). In the future, HIA:TTA and LDA:TTA have the potential to be used in a pre‐emptive manner to monitor the health of the HP, facilitate early diagnosis of diseases and study the pathophysiology of the organ.     

Effects of dietary supplementation with β‐glucan and synbiotics on growth,haemolymph chemistry,and intestinal microbiota and morphology in the Pacific white shrimp

The goal of this study was to investigate the effects of dietary supplementation with β‐glucan and microencapsulated probiotics (Bacillus subtilis or Pediococcus acidilactici) on growth performance, body composition, haemolymph constituents, and intestinal morphology and microbiota of the Pacific white shrimp Litopenaeus vannamei. Four treatment diets [basal diet (C), β‐glucan‐containing diet (β‐glu), β‐glucan plus B. subtilis‐containing diet (β‐glu+Bs), and β‐glucan plus P. acidilactici‐containing diet (β‐glu+Pa)] were fed to L. vannamei for 90 days. Shrimp fed the β‐glu and β‐glu+Pa diets exhibited similar growth performance and body protein content, which were significantly higher than those of shrimp fed the control diet (P < 0.05). No significant differences in haemolymph triglyceride, cholesterol, protein, haemolymph urea nitrogen or chloride were detected among the experimental diets. However, dietary β‐glucan alone increased the haemolymph glucose level and osmolarity (P < 0.05). Synbiotic supplementation had greater effects on intestinal microbiota and morphology than dietary β‐glucan alone. For example, β‐glu+Bs increased the number of intestinal lactic acid bacteria and decreased the number of Vibrio spp. (P < 0.05), and β‐glu+Pa increased the height of intestinal villi.     

Determination of poly‐β‐hydroxybutyrate assimilation by postlarval whiteleg shrimp,Litopenaeus vannamei using stable 13C isotope tracing

A preliminary study was conducted to demonstrate fate of the bacterial storage compound poly‐β‐hydroxybutyrate (PHB) once ingested and degraded in vivo in crustaceans. The 2% supplementation of ¹³C‐labeled Ralstonia eutropha DSM545 containing 75% PHB on dry weight in postlarval whiteleg shrimp, Litopenaeus vannamei resulted in consistent enrichment of two fatty acids (containing 14–24 carbons [C] in length ) pentadecanoic acid (15:0) and palmitic acid (16:0) in all lipid fractions. This could indicate that ¹³C signal may not only originate from the PHB but partly from the structural components of the bacterial cell. There was a trend of lower ¹³C enrichment in various lipid fractions of the postlarvae as compared to the diet. These findings may have important implications as to potential of PHB as energy source rather than as building block. There was a rapid transfer of ¹³C in the postlarvae as seen by the significant ¹³C enrichment in the tissues 4 hr after feeding with labeled bacterial cells. Overall, the assimilation of PHB in crustacean tissues is demonstrated for the first time. Our results indicate that PHB once ingested is rapidly assimilated in the tissues and could probably act as an energy source rather than as a building block.     

Effects of starvation on survival,growth and development of Exopalaemon carinicauda larvae

This study aimed at evaluating the effects of two starvation modes (fasting‐feeding mode and feeding‐fasting mode) on survival, growth and development of mysis larvae of Exopalaemon carinicauda. It showed that food restriction significantly affected the survival, growth and development in both starvation modes. With the extension of initial starvation period, a significantly reduced survival rate during metamorphosis was observed. In the fasting‐feeding mode, the duration of metamorphosis was extended and the body lengths of the individuals, which experienced successful metamorphosis to the post‐larval stage 1, were much shorter. In the feeding‐fasting mode, the extension of initial feeding period led to dramatically increased survival rate and body lengths, while there was no significant difference in duration of metamorphosis of those groups that reached to post‐larval stage. The 50% point‐of‐no‐return and the 50% point‐of‐reserve‐saturation were 3.85 days and 4.81 days respectively. The results of this study indicate that E. carinicauda mysis have the ability to withstand or recover from relatively extended starvation, but to ensure the success of commercial seedling, timely and adequate feeding is necessary. The information obtained from this study could provide a basis to optimize the feeding schedule of artificial seedling of E. carinicauda.     

Influence of temperature and Artemia enriched with ω‐3 PUFAs on the early ontogenesis of Atlantic sturgeon,Acipenser oxyrinchus Mitchill, 1815

Successful breeding of fish species in aquaculture depends on several factors, among which the temperature and feed are the most significant ones. This study was designed to evaluate the effect of temperature in the range of 14–18°C on the rate of embryogenesis and duration of larva period and to estimate the efficiency of Artemia nauplii enriched with PUFAs in growing sturgeon larvae. The temperature of 16°C is the most suitable for both egg incubation and Atlantic sturgeon prelarvae maintenance under aquaculture conditions. Even minor temperature fluctuations of 1 degree up or down leads to increased loss both of eggs and prelarvae. Increased temperature shortens the incubation period but has a lesser impact on the duration of prelarvae onset of external feeding. The technology of Artemia nauplii bioencapsulation with a PUFA‐containing supplement in A. oxyrinchus rearing increases sturgeon larvae weight by 1.5 times at a constant survival dynamic.     

Effects of continuous and alternate administration of β‐glucan and mannan‐oligosaccharide on the growth,immunity and resistance against Vibrio splendidus of sea cucumber Apostichopus japonicus (Selenka)

A 4‐week growth trial was conducted to compare the effects of different feeding strategies of dietary immunostimulants on the growth, immunity and resistance against Vibrio splendidus of sea cucumbers Apostichopus japonicus (Selenka). Six feeding strategies were set, including feeding immunostimulants‐free diet continuously (control), feeding dietary β‐glucan (1.25 g kg^?1 diet) continuously, feeding dietary mannan‐oligosaccharides (MOS; 2.00 g kg^?1 diet) continuously, feeding β‐glucan 2 days followed by MOS 5 days alternately, feeding β‐glucan 5 days followed by MOS 2 days alternately and feeding β‐glucan 7 days followed by MOS 7 days alternately. The sea cucumbers fed immunostimulants showed higher specific growth rate (SGR) and lower cumulative mortality than control (P < 0.05). When sea cucumbers were fed with β‐glucan continuously, total coelomocytes counts and superoxide anion were significantly higher than control on the 4th day (P < 0.05). However, these two immune parameters were not significantly higher than those in control after the 18th day (P > 0.05). While sea cucumbers continuously fed MOS, these two immune parameters were not significantly higher than control until the 15th day. All immune parameters of the sea cucumbers fed with β‐glucan and MOS alternately were significantly higher than those in control during the experiment (P < 0.05). The sea cucumbers fed with β‐glucan 7 days followed MOS 7 days alternately showed the highest SGR and second lowest cumulative mortality. It was suggested that this feeding strategy is most suitable for sea cucumbers.     

Comparative effect of Freund's adjuvant and Aloe vera L. gel on the expression of TNF‐α and IL‐1β in vaccinated Cyprinus carpio L. with Aeromonas hydrophila C. bacterin

The comparative effects of Freund's and Aloe vera gel as adjuvants on the expression of IL‐1β and TNF‐α genes were studied in vaccinated common carp (Cyprinus carpio) with Aeromonas hydrophila bacterin. Fishes were intraperitoneally immunized with A. hydrophila bacterin in combination with Aloe vera gel or Freund's and also without any adjuvant. At day 28 after immunization, all groups were challenged by lethal dose of A. hydrophila (10⁷ cells/fish). Changes in the expression of IL‐1β and TNF‐α genes were evaluated in anterior kidney before challenge and 12, 24, 72 and 7 days postchallenge using quantitative real‐time PCR. Higher expression levels of both genes were observed in all vaccinated groups compared with non‐immunized group. Fishes which received Aloe vera gel showed higher expression of IL‐1β and TNF‐α in relation to animals which vaccinated with or without Freund's adjuvant. We concluded that Aloe vera gel in compared with Freund's adjuvant had a more stimulatory effect on the expression of immune‐related genes in vaccinated common carp and it can use as a novel adjuvant in aquaculture.     

Unveiling the influence of the environment on the migration pattern of the Atlantic pomfret (Brama brama) in North‐eastern Atlantic waters

Hydroclimatic variability is one of the main factors that drives inter‐annual changes in fish migration patterns. This study analyses the relationship between climate‐oceanographic factors and migration of the Atlantic pomfret (Brama brama) in NE Atlantic waters. Geo‐referenced catch data from logbooks of longliners operating in European Atlantic waters from 2002 to 2013 were linked to environmental indices at different temporal and spatial scales. Our results point to a strong influence of temperature at 200 m depth as the key factor along with the upwelling in the Galician (NW Iberian) waters. However, sea surface temperature (SST) indirectly affects the geographical display of Atlantic pomfret migration, and large migrations are observed in scenarios of high SSTs in the migratory area (c. above 14.7°C). Migrations are constrained during years when temperatures are below this threshold. A longer time‐series of annual landings (1950–2013) supports this evidence and highlights the significant influence of temperature at 200‐m depth along with the landings of the previous year. Length frequency distributions suggest an increase in size between consecutive seasons supporting the hypothesis that migration is a feeding strategy and a return to tropical waters of origin for spawning. Our study shows that the temperature of intermediate waters is a key variable in determining the northward migration of the Atlantic pomfret whereas density‐dependence and surface climatic conditions trigger secondary effects on the migration pattern of this species.     

Effect of starvation on survival and biochemical profile of newborn juvenile lined seahorses,Hippocampus erectus (Perry, 1810)

This study assessed the effect of starvation on survival and nutritional status of newborn juveniles H. erectus (<10 days) to optimize rearing protocols, thereby helping to reduce wildlife exploitation. Maximum starvation time (MST) was estimated through the survival of juveniles continuously starved from birth. Resistance to starvation and the effect of food re‐introduction after 1, 2, 4 and 6 days of starvation on survival and metabolite concentrations (total proteins, total lipids, acylglycerides, cholesterol, glucose) were also determined. Survival amongst continuously starved animals decreased from 6.6 ± 0.5 to 0% from days 9 to 10 of starvation. Seahorses under different starvation–refeeding treatments all had 100% survival up to day 5 of experiments. After 10 days, however, a 4‐day starvation period followed by refeeding showed negative effects with <50% survival. During continuous starvation, lipids were the first energy reserve used to maintain basal metabolism, followed by proteins. Except for cholesterol, all metabolite concentrations differed between continuous starvations and feeding. Despite high seahorse survival after 5 days in the absence of food, the recovery of the metabolic status is possible after a starvation period of no more than 2 days, since irreversible physiological changes compromising the ultimate survival of the organisms take place after this time.     

Effects of β‐conglycinin on growth performance,antioxidant capacity and intestinal health in juvenile golden crucian carp,Carassius auratus

The objective of this study was conducted to research the effects of β‐conglycinin in the diets on the growth performance, immunity function, antioxidant capacity and intestinal health of juvenile golden crucian carp (Carassius auratus). Five diets contained respectively (0, 20, 40, 60 and 80 g/kg) β‐conglycinin, and were used to feed juvenile golden crucian carp for 56 days. Final weight, weight gain and specific growth rate were significantly reduced by dietary β‐conglycinin (20–80 g/kg). Feed efficiency and protein efficiency were significantly reduced by dietary β‐conglycinin (40–80 g/kg). In hepatopancreas, the activities of T‐SOD, ACP, ALT and T‐AOC were significantly suppressed by dietary β‐conglycinin (20–80 g/kg). The activities of LZM, AKP, CAT and GPx were significantly reduced by dietary β‐conglycinin (40–80 g/kg). The activities of protease were significantly reduced and the content of MDA was significantly increased by dietary β‐conglycinin (60–80 g/kg). In proximal intestines, the activities of protease and CAT were significantly decreased by dietary β‐conglycinin (40–80 g/kg). In mid and distal intestines, the activities of protease and CAT were significantly inhibited by dietary β‐conglycinin (20–80 g/kg). In intestines, T‐AOC and GPx were significantly declined by dietary β‐conglycinin (20–80 g/kg). In proximal and mid intestines, the content of MDA were significantly increased by dietary β‐conglycinin (40–80 g/kg). In distal intestines, the content of MDA was significantly increased by dietary β‐conglycinin (20–80 g/kg). The expression of IGF‐I was significantly decreased and the expression of IL‐1β and TNF‐α was significantly increased by dietary β‐conglycinin (20–80 g/kg). The structural integrity of intestinal tissues were damaged by dietary β‐conglycinin (20–80 g/kg), the part of intestinal villus were shed, the part of epithelial cells were separated from lamina propria. Ultimately, these results suggested dietary β‐conglycinin should be <20 g/kg in formula feed of golden crucian carp.     

Effects of γ‐aminobutyric acid supplementation on the growth performance,serum biochemical indices and antioxidant status of pharaoh cuttlefish,Sepia pharaonis

This study evaluated the effects of dietary γ‐aminobutyric acid (GABA) on the growth performance, serum biochemical indices and antioxidant status of pharaoh cuttlefish, Sepia pharaonis. Cuttlefish were cultured in open‐culturing cement pool systems for 8 weeks. Six practical diets supplemented with graded levels of GABA (0, 20, 40, 60, 80 and 100 mg/kg) were formulated. Each diet was randomly assigned to triplicate groups of 60 cuttlefish (mean weight: 10.33 g), the cuttlefish were fed two times per day to apparent satiation. The results showed that the specific growth rate (SGR), weight gain (WG) and feed efficiency (FE) significantly increased with dietary GABA supplementation (p < .05). The survival rate (SR) and protein content in muscle significantly increased when 58.9 mg/kg GABA supplied. Moreover, the nitric oxide (NO) content and acid phosphatase (ACP) activity in serum were significantly increased with dietary GABA supplementation (p < .05), while the activity of aspartate aminotransferase (AST) in serum decreased significantly when supplied with GABA at 58.9 mg/kg (p < .05). In addition, dietary GABA improved antioxidation activity by significantly increasing the activities of superoxide dismutase (SOD) and catalase (CAT) but decreasing malondialdehyde (MDA) levels in the liver and gill (p < .05). On the basis of the quadratic regression analysis of FE, the optimum content of dietary GABA in S. pharaonis was estimated to be 55.3 mg/kg. The findings of this study demonstrated that dietary GABA had a positive effect on the growth performance, serum biochemical indices and antioxidant status of S. pharaonis.     

Reconstruction of Atlantic herring (Clupea harengus) recruitment in the North Sea for the past 455 years based on the δ13C from annual shell increments of the ocean quahog (Arctica islandica)

Understanding the recruitment variability of the Atlantic herring North Sea stock remains a key objective of stock assessment and management. Although many efforts have been undertaken linking climatic and stock dynamic factors to herring recruitment, no major attempt has been made to estimate recruitment levels before the 20th century. Here, we present a novel annually resolved, absolutely dated herring recruitment reconstruction, derived from stable carbon isotope geochemistry (δ¹³C), from ocean quahog shells from the Fladen Ground (northern North Sea). Our age model is based on a growth increment chronology obtained from fourteen shells. Ten of these were micromilled at annual resolution for δ¹³C analysis. Our results indicate that the anthropogenically driven relative depletion of ¹³C, the oceanic Suess effect (oSE), became evident in the northern North Sea in the 1850s. We calculated a regression line between the oSE‐detrended δ¹³C results (δ¹³C?) and diatom abundance in the North Sea, the regression being mediated by the effect of phytoplankton on the δ¹³C of the ambient dissolved inorganic carbon. We used this regression to build an equation mediated by a nutritional link to reconstruct herring recruitment using δ¹³C?. The reconstruction suggests that there were five extended episodes of low‐recruitment levels before the 20th century. These results are supported by measured recruitment estimates and historical fish catch and export documentation. This work demonstrates that molluscan sclerochronological records can contribute to the investigation of ecological baselines and ecosystem functioning impacted by anthropogenic activity with implications for conservation and stock management.     

Effects of α‐lipoic acid on growth performance,body composition,antioxidant profile and lipid metabolism of the GIFT tilapia (Oreochromis niloticus) fed high‐fat diets

The wide use of lipid as a non‐protein energy substitute has led to lipid metabolic problems in cultured tilapia. Therefore, studies that reduce the effects of high‐fat diets in genetically improved farmed tilapia (GIFT) are required. This study evaluated the optimum level and effects of dietary α‐lipoic acid (α‐LA) on growth performance, body composition, antioxidant capacity and lipid metabolism of GIFT tilapia. The basal diet (120 g/kg lipid) was supplemented with six concentrations of α‐LA at 0 (control), L300, L600, L900, L1200 and L2400 mg/kg diet to make the experimental diets, which were fed to GIFT tilapia juveniles (initial body weight: 0.48 ± 0.01 g) for 8 weeks. The weight gain of fish improved significantly in the L300 than other dietary treatments. The intraperitoneal fat index and lipid content of fish fed on the L2400 diet decreased significantly than those fed on the control diet. The activities of superoxide dismutase and glutathione peroxidase (GSH‐Px) in serum and liver were significantly higher in fish fed on the L300 diet than the control. The reduced GSH content of fish fed on the L300 in serum and liver was significantly higher than those fed on control diet. The malondialdehyde content in serum and liver was significantly lower in L300 than in the control. The adipose triglyceride lipase gene was significantly up‐regulated in fish fed on the L2400, but the diacylglycerol acyltransferase 2 gene was down‐regulated in adipose. The liver‐type fatty acid‐binding protein gene in the liver was significantly up‐regulated in fish fed on the L300 and L600 diets. Moreover, the acyl‐coenzyme A oxidase gene in liver was significantly up‐regulated in fish fed on the L300, L600, L900 and L1200 diets. Polynomial regression analysis indicated that 439–528 mg/kg α‐LA is an appropriate dosage in high‐fat diet to improve growth performance and relieve lipid oxidative damage by accelerating lipid catabolism and reducing lipid synthesis in GIFT tilapia.     

Effect of cultivated copepods (Acartia tonsa) in first‐feeding of Atlantic cod (Gadus morhua) and ballan wrasse (Labrus bergylta) larvae

The aim of this study was to compare the nutritional composition and effects of short periods with cultivated copepod nauplii versus rotifers in first‐feeding. Atlantic cod (Gadus morhua) and ballan wrasse (Labrus bergylta) larvae were given four different dietary regimes in the earliest start‐feeding period. One group was fed the copepod Acartia tonsa nauplii (Cop), a second fed enriched rotifers (RotMG), a third fed unenriched rotifers (RotChl) and a fourth copepods for the seven first days of feeding and enriched rotifers the rest of the period (Cop7). Cod larvae were fed Artemia sp. between 20 and 40 dph (days posthatching), and ballan wrasse between 36 and 40 dph, with weaning to a formulated diet thereafter. In addition to assessing growth and survival, response to handling stress was measured. This study showed that even short periods of feeding with cultivated copepod nauplii (7 days) had positive long‐term effects on the growth and viability of the fish larvae. At the end of both studies (60 days posthatching), fish larvae fed copepods showed higher survival, better growth and viability than larvae fed rotifers. This underlines the importance of early larval nutrition.     

The primary culture and subculture of lymphoid cells from shrimp,Penaeus chinensis on thermo‐sensitive CS/α, β‐GP hydrogel

A thermo‐sensitive hydrogel composed of chitosan and α, β‐glycerophosphate was successfully synthesized and used as basic substrate for primary culture and subculture of lymphoid cells from shrimp, Penaeus chinensis. When the ambient temperature increases from 4°C to 25°C, the chitosan/α, β‐glycerophosphate solution transforms into a semi‐solid three‐dimensional hydrogel with a rough surface. Primary lymphoid cells formed were round and grew homogenously over the surface of the hydrogel. The peroxidase activity indicated that cells seeded on the hydrogel suffered less stress than cells suspended in control culture flasks. The higher relative viability and alkaline phosphatase activity of primary cultured lymphoid cells indicated that the chitosan/α, β‐glycerophosphate hydrogel enhanced the growth and proliferation of lymphoid cells. Subsequent subculture of lymphoid cells starting from crumbled primary hydrogel culture was also successful.     

Response of the intestinal mucosal barrier of carp (Cyprinus carpio) to a bacterial challenge by Aeromonas hydrophila intubation after feeding with β‐1,3/1,6‐glucan

The effect of dietary β‐glucan on the bacterial community in the gut of common carp (Cyprinus carpio) was examined after oral application of Aeromonas hydrophila. Carp received either feed supplemented with 1% MacroGard^®, a β‐1,3/1,6‐glucan, or a β‐glucan‐free diet. Fourteen days after feeding, half of the carp from each group were intubated with 10⁹ colony‐forming units (CFU) of a pathogenic strain of A. hydrophila. Gut samples were taken 12 hr to 7 days after application and analysed using microbiological and molecular biological techniques (NGS, RT‐PCR‐DGGE). The reaction of the mucosa and the microbiota to an A. hydrophila intubation differed in carp fed with β‐glucan compared to carp from the control group. In β‐glucan fed carp, the total bacterial amount was lower but the number of bacterial species was higher. Bacterial composition was different for carp from both treatment groups. The number of mucin filled goblet cells was reduced in carp fed the β‐glucan diet. Mucus was obviously released from the goblet cells and was probably washed out of the gut together with high numbers of bacteria. This might be protective against pathogenic bacteria and, therefore, feeding with β‐glucan may provide protection against infections of the gut in carp.