Resistance of genetically different common carp, Cyprinus carpio L., families against experimental bacterial challenge with Aeromonas hydrophila

The objective of this study was to determine the differences in disease resistance against artificial infection with Aeromonas hydrophila between genetically different common carp families. Four strains differing in their origin and breeding history were selected from the live gene bank of common carp maintained at the Research Institute for Fisheries, Aquaculture and Irrigation (HAKI, Szarvas, Hungary) to establish families with wide genetic background: Szarvas 15 (15), an inbred mirror line; Tata (T) scaly noble carp; Duna (D), a Hungarian wild carp and Amur (A), an East Asian wild carp. A diallele mating structure was used to allow the assessment of genetic variation within and between the tested 96 families for a variety of traits. The existing technologies of fertilization and incubation of carp eggs, as well as larval and fingerling rearing had been modified because of the large number of baseline populations. Two challenge trials of the 96 families of carp with Aeromonas hydrophila were done. The 10 most resistant and 10 most susceptible families to A. hydrophila were identified from these two challenges. The crosses that produced the most resistant families were mainly those having parents from Tata and Szarvas 15 domesticated strains, while the most susceptible families were from the wild strains Duna and Amur.     

Extracts of Oliviera decumbens and Satureja khuzestanica as immunostimulants affect some innate immunity indices of Cyprinus carpio against Aeromonas hydrophila infection

In this study, the effect of extracts of two herbs (Oliviera decumbens and Satureja khuzestanica) on immune response of carp was investigated. At the beginning of experiment, fish were divided to two groups including vaccinated (using vaccine developed against A. hydrophila) and non‐vaccinated. Fish in both groups were fed diets containing O. decumbens and S. khuzestanica and combination of two herbal extracts for 5 weeks. Control fish (negative control) and fish vaccinated only (positive control) were fed basal diets without supplements of herbal extracts. Lysozyme activity, antibody titre, complement activity and bactericidal activity in serum were measured. After 5 weeks feeding, fish were infected with A. hydrophila and mortalities were recorded. In both experimental groups, no significant differences were found in terms of alternative haemolytic complement (ACH50) activity and antibody titres of Serum. In non‐vaccinated fish group, lysozyme and bactericidal activity of fish fed S. khuzestanica or combination of O. decumbens and S. khuzestanica was higher compared with control and other experimental treatments. In vaccinated fish group, the lysozyme and bactericidal activity was not significant in all treatments compared with control group. The results of this study showed that feeding non‐vaccinated and vaccinated carp with only S. khuzestanica or in combination with O. decumbens enhance only some immunity indices including lysozyme and bactericidal activity in non‐vaccinated fish and these extracts have no immunological stimulatory role on vaccinated individuals.     

Non‐specific immune responses and intestinal immunity of common carp (Cyprinus carpio) fed Jujube (Ziziphus jujube) fruit extract

This study investigated the effects of Jujube (Ziziphus jujube) fruit extract (JFE) as an excellent source of nutrients and phytochemicals on the innate immune responses and expression of genes involved in intestinal immunity of common carp (Cyprinus carpio) fingerlings. Three hundred healthy carps (10.78 ± 0.05 g) fingerlings were randomly allocated to 12 fibre glass tanks (300L; 15 fish per tank) and fed for 8 weeks with experimental diets which contained different levels of JFE (0%, 0.25%, 0.5%, or 1%). Thereafter, serum non‐specific immune parameters [total immunoglobulin (Ig) as well as alternative complement (ACH50) and lysozyme activity (SL)] and expression of genes involved in intestinal immunity (lyz, tnf‐alpha, il1b, il8, il10, and tgfb) were measured. The results revealed a significant elevation of Ig level in the JFE‐fed fish (p < 0.05) over the control fish. However, there was no significant difference among treated groups in case of serum total Ig. The highest levels of SL and ACH50 were observed in fish fed 1% of JFE. Nonetheless, no significant difference was noticed regarding SL and ACH50 of control and other supplemented groups (0.25% and 0.5% JFE). Also, expression of lyz, tnf‐alpha, and il1b genes increased significantly (p < 0.05) in JFE‐fed fish, in a dose‐dependent manner. On the other hand, significant (p < 0.05) down‐regulation of il110 and tgfb were noticed in treated groups. The present findings suggested that extraction of Jujube, Z. jujube fruit, possesses beneficial effects on immune responses of common carp.     

Protective effects of basil (Ocimum basilicum) ethanolic extract supplementation diets against experimental Aeromonas hydrophila infection in common carp (Cyprinus carpio)

This study evaluated the effects of basil (Ocimum basilicum) leaves extract supplement on growth, blood parameters and resistance to Aeromonas hydrophila in Cyprinus carpio fingerlings. Basil leaves were mixed thoroughly with feed at 0, 100, 200, 400, 800 and 1600 mg kg^?1 of diet and were fed in triplicate group for 2 months. At the end of this period, growth performance and survival were determined. Then fish were challenged intraperitoneally with A. hydrophila and mortalities were recorded up to 10 days post challenge. Results indicated that WBC, RBC, Ht, Hb, total protein, albumin and globulin were higher (P < 0.05) in fish fed diets containing O. basilicum compared with the control. Highest specific growth rate values were observed in 400 mg kg^?1 group. Feed conversion ratio was significantly lower (P < 0.05) in fingerlings fed 400 and 800 mg O. basilicum kg^?1 dry diet. After 10 days post challenge, total protein, WBC, RBC and Hb were significantly higher in 400 mg kg^?1 compared with other groups. These results reveal that a dietary O. basilicum leaves extract of 400 mg kg^?1 fed for 60 days leads to increased growth performance and survival rate as well as improved feeding efficiency in common carp fingerlings rendering them more resistant against infection by A. hydrophila.     

Enhancement of non‐specific immune responses in common carp,Cyprinus carpio,by dietary carotenoids obtained from shrimp exoskeleton

The immunostimulatory role of carotenoid extract from shrimp processing discards was evaluated by feeding common carp fingerlings with a diet containing carotenoid extract for 21 weeks at 100 and 200 mg kg^?1 astaxanthin levels. Haemoglobin content was significantly (P < 0.05) higher in the blood of fish fed with carotenoid diet (>8.1 g dL^?1) compared with that from fish fed with carotenoid deficient diet (6.86 g dL^?1) and also leukocyte counts were higher (P < 0.05). No differences (P > 0.05) were observed in total serum protein, globulin level and albumin‐globulin ratio, but albumin content was higher (P < 0.05). Respiratory burst activity was significantly (P < 0.05) higher, the serum lysozyme activity almost doubled and the serum bactericidal activity was significantly increased when the fish were fed with diet containing 200 mg kg^?1 of astaxanthin but no significant differences were observed in serum trypsin inhibitory activity. There was a significant (P < 0.05) increase in leukocyte myeloperoxidase activity due to dietary carotenoids. Challenging fish with Aeromonas hydrophila after the feeding period resulted in 50% mortality in the control group while in the group fed with diet containing 100 mg kg^?1 astaxanthin, the mortality rate was 15%. No mortality and even symptoms of infection was not observed in the group fed with diet containing 200 mg kg^?1 of astaxanthin. The study indicated that carotenoid extract from shrimp processing discards can effectively be used as immunostimulants in aquaculture of carps and dietary carotenoids were found to enhance various immune defence mechanisms and also provide protection against the infection of pathogen A. hydrophila.     

Effect of dietary willow herb,Epilobium hirsutum extract on growth performance,body composition,haematological parameters and Aeromonas hydrophila challenge on common carp,Cyprinus carpio

The present study was conducted to evaluate the use of Epilobium hirsutum extract in diet of common carp, Cyprinus carpio (20 ± 2 g). Different levels of plant extract (0%, 0.5%, 1, 3% and 3%+2‰ multivitamin (2 g of multivitamin per kg diet) were spread on commercial diet. The feed was offered by 8 weeks. Results showed that fish fed experimental diets had no significant difference (P>0.05) in specific growth rates, condition factor, feed conversion ratios and survival compared with control. There were no significant difference (P>0.05) in moisture, crude protein, crude lipid and ash content of common carp fed diets containing various levels of plant extract. The mortality of fish challenged by Aeromonas hydrophila have been recorded for 30 days after challenging, results showed that mortality decreased significantly (P<0.05) in fish fed diet containing plant extract compared with the control. The lowest mortality observed in group fed diet containing 3% plant extract. Haematological parameters showed that white blood cells had significantly (P<0.05) increased in infected and uninfected groups compared with the control. Red blood cells, haemoglobin and haematocrit had no significant change (P>0.05) in infected and uninfected groups compared with the control.     

Comparative study on immunomodulatory and growth enhancing effects of three prebiotics (galactooligosaccharide,fructooligosaccharide and inulin) in common carp (Cyprinus carpio)

This study investigated the effects of different prebiotics, including galactooligosaccharide (GOS), fructooligosaccharide (FOS) and inulin (INL), on skin mucosal immune parameters, humoral immune responses as well as performance of common carp (Cyprinus carpio). Two hundred and forty specimens (13.85 ± 0.85 g) were stocked in 12 fibreglass tanks assigned into three treatments and a control group. The experimental diets were formulated to have equal level (2%) of the prebiotics. At the end of the feeding trial, the highest skin mucus lysozyme activities and total immunoglobulin (total Ig) were observed in GOS‐fed group (P < 0.05). However, skin mucus protease activity showed no significant difference among different dietary groups (P < 0.05). Blood respiratory burst activity was significantly increased in all prebiotic‐fed fish compared to the control group (P < 0.05); the highest activity was observed in GOS treatment. Furthermore, evaluation of humoral immune response revealed that feeding with GOS‐supplemented diet significantly increased lysozyme and alternative complement (ACH50) activity as well as total Ig compared to the control and other prebiotic groups. While no significant difference was observed between FOS and INL groups, common carps fed GOS‐supplemented displayed improved (P < 0.05) growth performance, including final weight, weight gain, specific growth rate (SGR) and feed conversion ratio (FCR), compared to the control treatment. These results revealed that different prebiotics modulate carp growth and immune response in different manner, and GOS seems to be the most suitable prebiotic.     

Recovery of Bacillus mycoides,B. pseudomycoides and Aeromonas hydrophila from common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) with gill disease

During a 3‐month period from June to the end of August 2016, ~5% mortalities were observed in a farm with rainbow trout (Oncorhynchus mykiss Walbaum) and one farm of common carp (Cyprinus carpio L.) in Bulgaria. The disease was manifested by gill ulcers/rot, asphyxiation and bloody ascites. Aeromonas hydrophila was isolated from the internal organs of all the diseased fish. Bacillus mycoides or B. pseudomycoides were recovered from the gill lesions on diseased carp and rainbow trout, respectively, with identification achieved by conventional phenotyping and by sequencing of the 16S rRNA gene. In vivo experiments confirmed that all three organisms were pathogenic to rainbow trout.     

Growth performance,haematology, antioxidant status,immune response and histology of common carp (Cyprinus carpio L.) fed biofloc grown on different carbon sources

A 10 weeks trial was performed to investigate how different carbon sources (sugar beet molasses: SBM+BFT, sugar: S+BFT, corn starch: CS+BFT) along with control affect welfare status of common carp (Cyprinus carpio L.) fingerlings in biofloc‐based tanks. Three hundred healthy fingerlings (22.5 ± 0.2 g) were randomly distributed in 12 tanks (70 L) at a density of 8.02 kg/m³ (25 fish/tank). The fish in BFT treatments fed only 75% feeding rate of control. At the end of the experiment no differences were seen between the groups in case of growth performance, but the fish reared in CS+BFT had a significant lower food conversion ratio compared with the others (p < .05). Different carbon sources did not affect on haematological parameters (p > .05). Total serum protein and antibody concentration differed in treatments, and the highest values were found in S+BFT and CS+BFT treatments (p < .05). No significant differences were observed in case of lysozyme, superoxide dismutase and complement activity in treatments (p > .05), whereas the fish in BFT treatments showed a significant higher total antioxidant capacity and lower glutathione peroxidase than the control (p < .05). Different carbon sources resulted in no change in goblet and kupffer cells in intestine and liver respectively. The highest relative percentage survival was obtained in the CS+BFT and S+BFT (50%) in comparison with SBM+BFT (20%) treatment. The results obtained in this experiment, suggest that corn starch improves immune response, diseases resistance and histology of digestive and respiratory systems in carp fingerlings when used as a carbon source in zero water exchange system.     

Effect of dietary soybean lecithin on growth parameters,digestive enzyme activity,antioxidative status and mucosal immune responses of common carp (Cyprinus carpio)

Common carp, Cyprinus carpio (13.54 ± 0.48 g), were fed soybean lecithin‐incorporated diets for 8 weeks. After the feeding trials, the different groups of fishes were assessed by survival, growth parameters, digestive enzymes, antioxidant enzyme activities, mucosal immune parameters, etc. The lecithin‐incorporated diets (10–30 g kg^–1) helped to increase the weight gain significantly (p < .05) compared with control groups; activity of digestive enzymes was also significantly (p < .05) higher compared with experimental groups. The antioxidant enzymes including catalase and serum superoxide dismutase were significantly (p < .05) higher in the C. carpio fed lecithin‐incorporated feed than the ones fed the control feed. The serum thiobarbituric acid‐reactive substances and serum glutathione peroxidase activity did not differ significantly (p > .05) between control and experimental groups. The positive correlation supported that antibacterial activity was significantly (p < .05) increased with increasing concentration of lecithin in the diets. Among the different mucosal immune parameters, lysozyme, protease and esterase activities were significantly (p < .05) increased with lecithin‐incorporated diets. The results of this study confirm that 20 and 30 g kg^–1 lecithin inclusion in diets helped to enhance the activities of digestive enzymes, antioxidant enzymes, antibacterial factors and mucosal immune parameters in C. carpio.     

Effects of glutamate in low‐phosphorus diets on growth performance,antioxidant enzyme activity,immune‐related gene expression and resistance to Aeromonas hydrophila of juvenile mirror carp (Cyprinus carpio)

This study investigated the effects of glutamate (Glu) in low‐phosphorus diets on growth performance, haematological indices, antioxidant enzyme activity, immune‐related gene expression and resistance to Aeromonas hydrophila in juvenile mirror carp (Cyprinus carpio) (5.07 ± 0.02 g). Fish were fed either graded levels of Glu (0 g/kg, 5 g/kg,  10 g/kg and 20 g/kg, named G0, G0.5, G1 and G2, respectively) in a low‐phosphorus diet (15 g/kg NaH₂PO_4, 0.49), or a normal phosphorus diet ( 20 g/kg NaH₂PO_4, 0.61) without added Glu (C), for 8 weeks. At the end of the feeding trial, the fish were challenged with A. hydrophila. Compared with G0 group, 10 g/kg and 20 g/kg Glu supplementation of the low‐phosphorus diet significantly improved the final weight, WGR, SGR and PER, and decreased FCR (p < .05). Glu supplementation of the low‐phosphorus diet significantly enhanced the T‐AOC, SOD activity and GSH content in intestine (p < .05). Glu supplementation significantly reduced MDA content in foregut and midgut and increased CAT activity in midgut and hindgut (p < .05). Regarding immune‐related gene expression, Glu supplementation significantly diminished the up‐regulation of intestinal TNF‐α, IL‐1β and IL‐8 mRNA levels induced by phosphorus deficiency (p < .05). The survival rate of the G1 group was significantly higher than that of the G0 group (p < .05). In conclusion, 10 g/kg Glu supplementation in low‐phosphorus diets can improve the growth performance, enhance the activity of intestinal antioxidant enzymes and strengthen the immune function of juvenile mirror carp.