Koi herpesvirus and carp edema virus threaten common carp aquaculture in Croatia

Common carp (Cyprinus carpio) is a very important fish species for warm-water aquaculture in Croatia. All Croatian carp farms are subjected to a surveillance programme for the presence of koi herpesvirus (KHV), causing a deadly disease called koi herpesvirus disease (KHVD). However, there is no surveillance for other viral pathogens of importance like carp edema virus (CEV), a causative agent of koi sleepy disease (KSD). During regular testing within the KHVD surveillance programme, we tested samples for CEV simultaneously. The screening indicated possible outbreaks of KHVD and KSD. During 2016, KHVD broke out in an isolated area and soon thereafter a KHV eradication programme was successfully performed. However, during 2018 and 2019, two additional mortality events occurred in lakes in the southern part of Croatia during the spring. Samples from both events tested positive for CEV. An epidemiological investigation confirmed the introduction of infected carps from an infected farm to one of the lakes. To prevent the spreading of CEV into open waters, it is of utmost importance to introduce CEV testing before fish movement or to perform regular testing of all carp farms in the country to determine CEV prevalence for the purpose of implementation of control measures.     

Effects of antimicrobial peptides on growth,morphology of foregut villi and related genes mRNA expression in the common carp (Cyprinus carpio)

A 60‐day feeding trial was conducted to examine the effects of different levels (0, 100, 200, 400 and 600 mg/kg) of antimicrobial peptides on growth, protease activity of foregut, the morphology of foregut villi and related genes mRNA expression level in the common carp (Cyprinus carpio). The results showed that the feed of antimicrobial peptides promote common carp growth, and the optimal dosage of antimicrobial peptides is 200–333 mg/kg in the common carp feed. The protease activity of 200 and 400 mg/kg groups were significantly higher than the control and other groups (p < 0.05). The foregut villus height with 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The crypt depth of 200 and 400 mg/kg antimicrobial peptide groups were significantly lower than control group (p < 0.05). The ratio of villus height and crypt depth of 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The ratio with 600 mg/kg group was significantly lower than the control group (p < 0.05). The IGF‐I gene expression level of 200 mg/kg and 400 mg/kg groups were significantly higher than the control group and 600 mg/kg group (p < 0.05). The IL‐1β gene expression level of 100 mg/kg and 200 mg/kg groups were significantly higher than the control group (p < 0.05). These results indicated up‐regulation of growth and immune related genes in antimicrobial peptides fed common carp. Correlation analysis showed that IGF‐I mRNA and IL‐1β mRNA were positively correlated with SGR. IL‐1β mRNA and FCR were significantly negative correlated. It indicated that growth and immune gene common regulated the growth of the carp under antimicrobial peptides intervention. In conclusion, antimicrobial peptides can improve growth and related genes mRNA expression in the common carp. Further studies using molecular biological technique or immunologic methods are required to conclude that antimicrobial peptides are beneficial in common carp.     

Effects of a highly purified fucoidan from Undaria pinnatifida on growth performance and intestine health status of gibel carp Carassius auratus gibelio

A six‐week feeding trial was conducted to determine the effects of different concentrations of fucoidan (1 g/kg, 10 g/kg and 30 g/kg; w/w) from Undaria pinnatifida on gibel carp (Carassius auratus gibelio). Our results demonstrated that 30 g/kg fucoidan significantly increased (p < .05) growth performance, intestinal digestive enzyme activities, acid phosphatase activity and immunoglobulin M content. Histological examinations revealed that gibel carp receiving 30 g/kg fucoidan had significant higher abundance of mucin‐containing goblet cells in middle and distal intestine as compared with control treatment (p < .05). Intestinal microbiota analysis showed that 30 g/kg fucoidan supplementation significantly increased (p < .05) the abundance of Cetobacterium and Aeromonas, but lowered (p < .05) the prevalence of pathogenic bacteria Plesiomonas and a mucin‐degrading bacterium Mucinivorans. Furthermore, RNA‐seq and RT‐qPCR analysis indicated that 30 g/kg fucoidan caused significant changes (p < .05) in the expression of genes involved in immune regulation (such as interleukin‐8 and cyclooxygenase), signal transduction (such as phosphatidylinositol‐4,5‐bisphosphate 3‐kinase and protein kinase B) and nutrition utilization (maltase–glucoamylase and muscarinic acetylcholine receptor 3). Together, the current study shows that fucoidan supplementation could elevate the activity of intestinal digestive enzymes, modulate intestinal microbial communities and potentiate a higher state of immune readiness, which might consequently improve growth performance and intestine health status of gibel carp.     

Dietary Moringa oleferia leaf meal induce growth,innate immunity and cytokine expression in grass carp,Ctenopharyngodon idella

The present research was designed to investigate the growth promoting and immunostimulating properties of Moringa oleferia leaf meal (MLM) in grass carp. Juvenile grass carp (22.03 g ± 1.164) were fed with diets supplemented with 0, 50, 100 and 150 g/kg MLM for 48 days. At the end of feeding trial, skin mucus was used for analysis of lysozyme, protease, antiprotease and peroxidase activity. Head kidney was used for expression analysis of tumour necrosis factor‐alpha, interleukin‐8 and interferon‐γ. The obtained results showed that fish fed with 100 and 150 g/kg MLM had significant increase in weight gain and specific growth rate (p < .05). However, condition factor was not altered. The MLM (50 and 100 g/kg) inclusion resulted in higher mucus lysozyme and protease activity (p < .05), while peroxidase activity increased only in fish fed with 100 g/kg MLM and antiprotease activity was not altered. Expression of tnf‐α increased in a dose‐dependent manner, and significant (p < .05) increase was recorded in fish fed with 150 g/kg MLM. The expression of il‐8 and ifn‐γ increased in fish fed with 50 and 150 g/kg MLM; however, the increase was not significant (p > .05). In conclusion, supplementing juvenile grass carp feed with MLM up to 150 g/kg has growth promoting and immunostimulating effects.     

Effect of substitution of dietary fish meal by soybean meal on different sizes of gibel carp (Carassius auratus gibelio): digestive enzyme gene expressions and activities,and intestinal and hepatic histology

A 7‐week growth trial was conducted to evaluate the effects of dietary soybean meal (SBM) on digestive enzyme activity of intestinal mucosa, mRNA levels of digestive enzymes in hepatopancreas, and the mid‐intestinal and hepatopancreas histology of gibel carp CAS III (Carassius auratus gibelio). Four different growth phases of gibel carp (initial body weight: fry, 0.8 g; juvenile, 5.0 g; 1‐year‐old, 62.7 g; and broodstock, 135.6 g) were tested. Seven isonitrogenous and iso‐energetic diets were formulated to contain different SBM replacement levels (0%, 20%, 40%, 60%, 80% and 100% of dietary fish meal protein), and another diet (SBMAA) contained all SBM protein and supplied crystalline amino acids. The results showed that the activities of mid‐intestine trypsin, α‐amylase and gamma glutamyl transpeptidase reduced with increased dietary SBM, while the chymotrypsin activity increased first and then decreased. The ultrastructures of intestinal epithelial cells and hepatopancreas cells in fry and broodstock fish were distinctly affected by 200 g kg^‐1 dietary SBM. Supplementation of dietary amino acid to the highest replacement groups was not sufficient to improve digestive and absorptive capacities and growth performance. Gibel carp may be adapted to dietary SBM through increase in gene expression of hepatopancreas digestive enzymes and has potential to utilize proceeded SBM as feedstuffs.     

三氯生对红白鲫运动行为的影响

利用视频跟踪系统研究三氯生(TCS)对红白鲫运动行为的影响。将红白鲫(red-white crucian carp)暴露在5种不同浓度的TCS(0.50、0.60、0.70、0.80和0.90 mg/L)水体中,视频记录5 min内红白鲫的运动轨迹(运动轨迹线、摆动次数)、游动速度和垂直穿梭次数的变化。同时,进行TCS(0.60、0.69 mg/L)对红白鲫的14 d暴露实验,暴露实验结束后,2 min内监测上述指标。结果显示:短期TCS暴露,红白鲫的运动轨迹条数、摆动次数、垂直穿梭次数、运动速率显著增加;暴露14 d后的红白鲫上述运动行为学指标显著降低。结果表明:在一定浓度范围内,TCS能够改变红白鲫的运动行为。     

The influence of diet on the grass carp intestinal microbiota and bile acids

To elucidate the influence of different diet on the intestinal microbe and bile acids, we characterized the microbiota and bile acids in the hindgut content of grass carp fed on formula feed (FF group) or Sudan grass (SG group). Fusobacteria and Proteobacteria were significantly more represented in FF group than in SG group whereas Bacteroidetes was significantly more abundant in SG group than in FF group. Simpson diversity was significantly higher in FF group than in SG group (t = 2.33, P < 0.05). Chenodeoxycholic acid (CDCA) was the most abundant primary bile acid in the two groups, with average concentrations of 1.03 ± 0.62 and 4.44 ± 1.80 ng mg^?1 in SG and FF group respectively. The most abundant secondary bile acid was deoxycholic acid (DCA) in SG group and ursodeoxycholic acid (UDCA) in FF group, with average concentrations of 0.17 ± 0.06 and 2.67 ± 0.88 ng mg^?1 respectively. UDCA is significantly more abundant in FF group than in SG group, and the total bile acids were higher in FF group than in SG group. Cetobacterium and Fusobacteriaceae U114 were significantly related with the concentrations of CDCA (r = 0.85, P < 0.05 and r = 0.82, P < 0.05 respectively) and UDCA (r = 0.92, P < 0.01 and r = 0.92, P < 0.01 respectively). However, Bacteroides was negatively related with the concentration of UDCA (r = ?0.67, P < 0.05). Overall, there existed certain relationship between the intestinal microbes and the faecal bile acids, and they were both influenced by the diet.     

Isolation of infectious microalga Prototheca wickerhamii from a carp (Cyprinus carpio) – a first confirmed case report of protothecosis in a fish

Protothecosis is a rare infection caused by environmentally ubiquitous achlorophyllic microalgae of the genus Prototheca. Here, we describe a first case of protothecosis in a carp (Cyprinus carpio), which is at the same time the first case of protothecosis in a fish, confirmed by phenotype‐ and molecular‐based methods, including PCR sequencing of the rDNA cluster and protein profiling using matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry.