Molecular characterizations, immune modulation, and antibacterial activity of interleukin-17A/F1a and interleukin-17A/F1b in common carp Cyprinus carpio

Interleukin (IL)-22 in common carp (Cyprinus carpio L.): Immune modulation, antibacterial defense, and activation of the JAK-STAT signaling pathway

Molecular characteristics of interleukin (IL)-17A/F3 and its immune response on the pathogen and functional regulation on cytokines in common carp Cyprinus carpio L.

Interleukin-15 (IL-15) is a crucial cytokine involved in immune system regulation, which is produced by various cell types, including dendritic cells, monocytes, and macrophages. IL-15 plays a key role in the proliferation and activation of natural killer (NK) cells, CD8+ T cells, and memory CD8+ T cells, supporting their survival and enhancing their effector functions. Although IL-15 homologues in fish have been identified, their functions remain poorly understood. In this study, we cloned and investigated the bioactivities of an IL-15 homologue, referred to as IL-15 like (CcIL-15L), in common carp (Cyprinus carpio L.). An expression pattern analysis revealed that CcIL-15L was constitutively expressed in all examined tissues of healthy common carp, with the highest expression level observed in the intestine. Additionally, CcIL-15L expression was significantly up-regulated in the head kidney, spleen, gills, and intestine following Aeromonas hydrophila infection. In vitro, the recombinant protein CcIL-15L can significantly up-regulated the gene expression levels of pro-inflammatory cytokines (IL-1β, IL-6, IFN-γ, and TNF-α) and NK cell activation (perforin and Eomesa). We constructed a 3×FLAG eukaryotic expression vector and successfully expressed it in common carp by intramuscular injection. Additionally, the heterologous CcIL-15L protein was successfully overexpressed in vivo, and immune-related genes including CD4-1, CD8β2, TNF-α, and IgM showed significant induction in the head kidney and spleen. Furthermore, CcIL-15L overexpression reduced the bacterial loads after 24 h post-A. hydrophila infection in the liver, spleen, and kidney. Phagocytic and chemotaxis assays showed that rCcIL-15L could promoted the phagocytosis and chemotactic abilities of common carp HKLs. Our study provides a new perspective on the role for CcIL-15L in immunological functions in common carp.

This research aims to know the role of maggot flour protein as a fishmeal protein source replacement on growth performance and survival of common carp (Cyprinus carpio) and the optimum dose from artificial maggot flour feed. This research take a place at Aquaculture Laboratorium of Diponegoro University in 23 January – 6 March 2023. The experimental method using complete randomized design (RAL) consisting of 4 treatments 3 repetitions. The treatment is applied with percentage maggot flour substitution 0%, 10%, 20%, and 30% per gram of feed. The substitution set according to carp protein sufficient (30 – 35%), so that carp does not having excess protein that can effect on growth performance. The test fish used was common carp with an average size of 4.54 ± 0.23 cm with an average weight of 0.75 ± 0.03 gr/head. The experimental containers using a aquarium tank with size of 45cm x 30cm x 22.5cm. Stocking density per aquarium is 0.5 fish/liter with a maintenance period of 42 days. The experimental using feeding methods 5% of biomass with a frequency 3 times/day. The experimental results showing that substitution of artificial maggot flour feed for common carp is significantly influential (P<0.05) on specific growth rate (SGR), feed conversion ratio (FCR), and survival rate (SR) of common carp. Based on experimental, the best dose substitution artificial maggot flour feed has been shown in treatments C (20%) and D (30%). Treatment C produced SGR 1.93 ± 0.04 and FCR 2.41 ± 0.07 and treatment D produced SGR 1.97 ± 0.09% and FCR 2.34 ± 0.12. The results of observations during the research showed that common carp fed with the experimental feed (substitution of maggot flour for artificial feed 0%, 10%, 20%, 30%) had a high survival rate with an average survival rate above 90%.

A 28-days trial was conducted in an aquaponics culture system to determine the effect of the different vegetable plants application on the growth performance and survival rate of common carp (Cyprinus carpio). Common carp seeds were obtained from local fish farmers in Aceh Besar District. A total of 360 common carp fingerlings (size 3-5 cm) were used and placed in 12 containers with the capacity of 80 L each. The study used four treatments, namely A (control), B Kale (Ipomoea aquatica), C Lettuce (Lactuca sativa), and D Pakcoy (Brassica rapa subsp. chinensis), with three replications for each treatment. The result showed that different vegetable plants' applications significantly affected (P 0.05) the absolute weight gain, absolute length gain, and survival rate of common carp fingerlings. The highest value obtained was in treatment B (Ipomoea aquatic) with an absolute weight gain of 3.93 ± 0.28 g, absolute length gain of 3.81±0.21 cm, and survival rate of 100±0.0%, while the lowest was in treatment A (control) with an absolute weight gain of 1.21±0.76 g, absolute length gain of 1.70±0.10 cm and survival rate of 91±0.86%. Water quality was also collected in this study and the parameters during rearing were still within the tolerance limits of common carp rearing. Therefore, it can be concluded that the Kale plant, Ipomoea aquatica is the most recommended vegetable plant to be co-cultured with common carp in the aquaponics system.Keywords:AquaponicsCommon CarpVegetable Plants

The purpose of this study was to determine the effect of adding phytase enzyme in the feed on digestibility of feed, efficiency of feed utilization, relative growth rate and survival rate of Common carp (Cyprinus carpio). Fish samples in this research were Common carp with an average - weight of 3.34 ± 0,16 g/fish. The treatments were adding the phytase enzyme in the feed with the different level of doses. Those were A (0 U kg-1 feed), B (500 U kg-1 feed), C (1.000 U kg-1 feed g) and D (1.500 U kg-1 feed). Observation was conducted on digestibility of protein (ADCP), digestibility of phosphor (ADCF), efficiency of feed utilization (EFU), relative growth rate (RGR), protein efficiency ratio (PER), feed conversion ratio (FCR), survival rate (SR) and water quality parameters. The results show that the addition of phytase enzyme significantly (P<0.01) affected on ADCP, ADCF, EFU, RGR, FCR, and PER, on the other hand it insignificantly (P>0.05) affected on SR of common carp. Based on results, it was concluded that optimum doses of phytase enzyme feed in terms of digestibility of feed, efficiency utilization of Feed and growth rate of Common carp ranges from 943 to 1100 U kg-1 feed

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89 ± 6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29 × 108 colony forming unit/mL (CFU/mL; LD50 dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P < 0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~ 39.94 and ~ 4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~ 5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P < 0.05) increase in total protein content of blood plasma (~ 43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.

This study was aimed to estimate the selectivity properties of multifilament trammel and gillnets for common carp ( Cyprinus carpio L., 1758) in Marmara Lake in Manisa, Turkey. Trammel and gillnets of 4, 6, 8, and 10 cm stretched mesh size and 210 d/2 no twine thickness were tested in the Marmara Lake. In addition, selectivity parameter of 11, 12, 13 and 14 cm mesh size of trammel and gillnets were modelled. SELECT method was used to estimate the selectivity parameters. Experiments were carried out monthly in the three different stations identified eastern, middle and western areas which represent longitudinal length of the lake. A total of 36 trials were conducted both nets. A total of 119 common carps were caught ranges between 11.3 - 49.0 cm total lengths. According to the Bi- normal model, which gave the lowest deviance for both trammel and gillnets, selectivity curves were estimated for 4, 6, 8, 10, 11, 12, 13 and 14 cm stretched mesh size modelled lengths as 12.98, 19.47, 25.96, 32.45, 35.70, 38.94, 42.19 and 45.43 cm for trammel nets, 12.40, 18.60, 24.80, 31.00, 34.10, 37.20, 40.30 and 43.40 cm for gillnets, respectively. When considering minimum landing size (40 cm total length), it is recommended that both nets under 13 cm mesh size should be prohibited for common carp fishery in the Lake Marmara. Keywords: Trammel net, gillnets, common carp Cyprinus carpio, size selectivity, Lake Marmara

The ATP-binding cassette (ABC) gene family is considered to be one of the largest gene families in all forms of prokaryotic and eukaryotic life. Although the ABC transporter genes have been annotated in some species, detailed information about the ABC superfamily and the evolutionary characterization of ABC genes in common carp (Cyprinus carpio) are still unclear. In this research, we identified 61 ABC transporter genes in the common carp genome. Phylogenetic analysis revealed that they could be classified into seven subfamilies, namely 11 ABCAs, six ABCBs, 19 ABCCs, eight ABCDs, two ABCEs, four ABCFs, and 11 ABCGs. Comparative analysis of the ABC genes in seven vertebrate species including common carp, showed that at least 10 common carp genes were retained from the third round of whole genome duplication, while 12 duplicated ABC genes may have come from the fourth round of whole genome duplication. Gene losses were also observed for 14 ABC genes. Expression profiles of the 61 ABC genes in six common carp tissues (brain, heart, spleen, kidney, intestine, and gill) revealed extensive functional divergence among the ABC genes. Different copies of some genes had tissue-specific expression patterns, which may indicate some gene function specialization. This study provides essential genomic resources for future studies in common carp.

Cloning and characterization of two duplicated interleukin-17A/F2 genes in common carp ( Cyprinus carpio L.): Transcripts expression and bioactivity of recombinant IL-17A/F2

Iron uptake, transport, and storage require the involvement of several proteins, including ferroportin (fpn), the sole known iron efflux transporter. Due to its critical function fpn has been studied, particularly in humans. Here, we characterized the ferroportin gene in common carp (Cyprinus carpio L.) and performed RNA-seq analysis to evaluate its constitutive transcription levels across different tissues. Our results indicate that C. carpio possesses two functional fpns with distinct expression patterns, highlighting the potential for functional divergence and expression differentiation among fpns in this species.

In humans, the frizzled (FZD) gene family encodes 10 homologous proteins that commonly localize to the plasma membrane. Besides being associated with three main signaling pathways for cell development, most FZDs have different physiological effects and are major determinants in the development process of vertebrates and. Here, we identified and annotated the FZD genes in the whole-genome of common carp (Cyprinus carpio), a teleost fish, and determined their phylogenetic relationships to FZDs in other vertebrates. Our analyses revealed extensive gene duplications in the common carp that have led to the 26 FZD genes that we detected in the common carp genome. All 26 FZD genes were assigned orthology to the 10 FZD genes of on-land vertebrates, with none of genes being specific to the fish lineage. We postulated that the expansion of the FZD gene family in common carp was the result of an additional whole genome duplication event and that the FZD gene family in other teleosts has been lost in their evolution history with the reason that the functions of genes are redundant and conservation. Through the expression profiling of FZD genes in common carp, we speculate that the ancestral gene was likely capable of performing all functions and was expressed broadly, while some descendant duplicate genes only performed partial functions and were specifically expressed at certain stages of development.

Swimming, grazing and social behaviour of rohu Labeo rohita (Hamilton) and common carp Cyprinus carpio (L.) in tanks under fed and non-fed conditions

Evaluation of probiotic characteristics and whole genome analysis of Bacillus velezensis R-71003 isolated from the intestine of common carp (Cyprinus carpio L.) for its use as a probiotic in aquaculture

To evaluate the functions of safflower on aquatic animal, the effects of dietary safflower supplementation on growth performance, immunity response, short-chain fatty acids, and intestinal microflora of common carp (Cyprinus carpio L.) were studied in the present study. Four isonitrogen and isolipid experimental diets were formulated by adding 0, 0.25%, 0.50%, and 1.00% safflower to the basal diet (CON, S-0.25%, S-0.50%, and S-1.00%, respectively). The results showed that safflower supplementation (S-0.50%) significantly improves the growth performance (FBW, WGR, SGR) compared with that of the CON group ( P &lt; 0.05 ). Similarly, safflower also promoted the CAT and LZM activities in the midgut, yet the content of MDA in the S-0.50% group was reduced ( P &lt; 0.05 ). The content of proinflammatory factors (IL-1β, IL-12) in serum underwent significant reduction ( P &lt; 0.05 ); the level of anti-inflammatory factor IL-10 had an upward trend in the S-0.50% group. Moreover, the mRNA expression levels of the IL-10, TGF-β, NF-κB, and TLR-1 gene in the head kidney were remarkably upregulated ( P &lt; 0.05 ) in safflower addition groups, while IL-6 and IL-1β were downregulated significantly in the S-1.00% group ( P &lt; 0.05 ). After 96 hours of the A. hydrophila challenge test, the survival rate of common carp had been improved greatly with the safflower addition, which was most impressive in the S-1.00% group. Additionally, the levels of acetic acid and butyric acid in intestinal contents were significantly higher in the S-0.50% and S-1.00% groups than in the CON group ( P &lt; 0.05 ). Dietary safflower increased the intestinal microbiota abundance of Fusobacteria at the phylum level. At the genus level, the abundance of beneficial bacteria (Lactobacillus and Bifidobacterium) increased significantly in the S-1.00% group; some pathogens such as Aeromonas, Shigella, and Streptococcus were inhibited. Spearman coefficient analysis revealed that the contents of SCFAs were positively correlated with probiotics and negatively correlated with pathogens. In summary, safflower addition at an appropriate dose can promote the immune functions of the common carp by producing the short-chain fatty acids and regulating the intestinal microflora.