ABSTRACT
Koi(Cyprinus carpio) is an ornamental variety of common carp frequently kept as pets. Given their long lifespan, neoplasia, albeit uncommon, may occur in these animals, and only a few studies have faced their cytological diagnosis. In the present case, a koi carp was referred to the clinicians due to coelomic swelling. The carp underwent surgery, which revealed an enlargement of both testes. Testicular samples were cytologically and histologically examined. The lesion was diagnosed as a seminoma since it was composed of round, large, atypical, and often multinucleated cells with round central nuclei and moderate cytoplasm. These tumors had the same appearance as seminomas in mammals and should be considered among differential diagnoses when coelomic swelling occurs in koi carp. Seminomas in koi carp are diagnosed histologically, but cytology, a rapid and cheap exam executable in all veterinary clinical facilities, could be a relevant preliminary diagnostic tool that may influence the entire diagnostic process.
Subject(s)
Carps , Seminoma , Testicular Neoplasms , Animals , Male , Seminoma/veterinary , Seminoma/diagnosis , Seminoma/pathology , Testicular Neoplasms/veterinary , Testicular Neoplasms/pathology , Testicular Neoplasms/diagnosis , Fish Diseases/diagnosis , Fish Diseases/pathology , Testis/pathology , Rapid Diagnostic Tests , CytologyABSTRACT
A rapidly growing nontuberculous mycobacterium was isolated from diseased koi carp in Niigata, Japan, which was identified as representing a novel Mycolicibacterium species through whole genome sequence analysis. The bacterial isolates (NGTWS0302, NGTWS1803T and NGTWSNA01) were found to belong to the genus Mycolicibacterium through phylogenetic analysis using whole genome sequences of mycobacteria species. The bacterial colony was smooth, moist and non-chromogenic on 1% Ogawa medium at 30â°C. In biochemical characteristic tests, the bacterial isolates showed positive reactions for catalase activity, Tween 80 hydrolysis and tellurite reduction. The isolates were sensitive to 2-4 µg ml-1 ampicillin, kanamycin and rifampicin. Based on these results, we propose a novel Mycolicibacterium species, Mycolicibacterium cyprinidarum sp. nov. The type strain is NGTWS1803T (=JCM 35117T=ATCC TSD-289T).
Subject(s)
Bacterial Typing Techniques , Carps , DNA, Bacterial , Phylogeny , RNA, Ribosomal, 16S , Animals , Carps/microbiology , Japan , DNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Fish Diseases/microbiology , Anti-Bacterial Agents/pharmacology , Fatty Acids , Microbial Sensitivity Tests , Whole Genome Sequencing , Base CompositionABSTRACT
Ulcerative dermatitis (UD) is common in ornamental fish collections and is typically associated with a wide range of bacterial aetiologies. Clinical reports describing Shewanella xiamenensis-associated UD are limited, however, despite growing attention to pathogenic Shewanella species in fish. Two out of 95 koi carp with UD were presented for clinical assessment by a commercial collection (n = 3000 fish) and subsequently killed on welfare grounds for necropsy. Both specimens exhibited extensive cutaneous ulcers and coelomic fat necrosis with petechial haemorrhages on post-mortem examination. Shewanella xiamenensis was cultured from ulcerated skin tissues taken from both fish, with consistent intralesional gram-negative rod-like bacteria seen on skin scrape cytology. Histology also confirmed intralesional gram-negative rod-like bacteria within multiple ulcerative and erosive dermatitis lesions, plus myofibre necrosis and necrotising coelomic steatitis, in both specimens. Features associated with impaired generalised osmoregulation secondary to UD were detected within the striated muscle underlying the ulcers, the gills, and the caudal aspects of the kidneys. Additional histological features suggestive of sepsis were also seen in one of the fish. In the interim period, morbidity had increased from 3.2% to around 30% of the entire stock. Following culture results, increased pond water changes were implemented (q.2-3d) and the remaining stock was treated with florfenicol, resulting in complete resolution of UD in the collection (as per client). This article highlights the first description of S. xiamenensis-associated UD in koi carp/diseased ornamental fish in the United Kingdom.
Subject(s)
Carps , Fish Diseases , Gram-Negative Bacterial Infections , Shewanella , Animals , Shewanella/isolation & purification , Fish Diseases/microbiology , Fish Diseases/pathology , Gram-Negative Bacterial Infections/veterinary , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/pathology , Skin Ulcer/veterinary , Skin Ulcer/microbiology , Skin Ulcer/pathology , Dermatitis/veterinary , Dermatitis/microbiology , Dermatitis/pathologyABSTRACT
Viperin, also known as radical S-Adenosyl methionine domain containing 2 (RSAD2), is an IFN stimulated protein that plays crucial roles in innate immunity. Here, we identified a viperin gene from the koi carp (Cyprinus carpio) (kVip). The ORF of kVip is 1047 bp in length, encoding a polypeptide of 348 amino acids with neither signal peptide nor transmembrane protein. The predicted molecular weight is 40.37 kDa and the isoelectric point is 7.7. Multiple sequence alignment indicated that putative kVip contains a radical SAM superfamily domain and a conserved C-terminal region. kVip was highly expressed in the skin and spleen of healthy koi carps, and significantly stimulated in both natural and artificial CEV-infected koi carps. In vitro immune stimulation analysis showed that both extracellular and intracellular poly (I: C) or poly (dA: dT) caused a significant increase in kVip expression of spleen cells. Furthermore, intraperitoneal injection of recombinant kVip (rkVip) not only reduced the CEV load in the gills, but also improved the survival of koi carps following CEV challenge. Additionally, rkVip administration effectively regulated inflammatory and anti-inflammatory cytokines (IL-6, IL-1ß, TNF-α, IL-10) and interferon-related molecules (cGAS, STING, MyD88, IFN-γ, IFN-α, IRF3 and IRF9). Collectively, kVip effectively responded to CEV infection and exerted antiviral function against CEV partially by regulation of inflammatory and interferon responses.
Subject(s)
Carps , Fish Diseases , Poxviridae Infections , Poxviridae , Animals , Carps/genetics , Edema , Interferons , Antiviral Agents/pharmacologyABSTRACT
In this study, we evaluated gamete quality parameters of mature male koi carp (Cyprinus carpio) exposed to three different concentrations (1, 10, and 100 µg/L) of di-(2-ethylhexyl) phthalate (DEHP). After 60 days of exposure, there was a significant decrease in the gonadosomatic index (GSI) of males exposed to 10 and 100 µg/L of DEHP. Histological analysis of the testes revealed impaired histoarchitecture, including inflammatory cells, intratubular vacuoles, and swollen seminiferous tubules in treatment groups. Gamete quality parameters like sperm production, motility, spermatocrit, and sperm density values were significantly decreased at the 10 and 100 µg/L concentrations. Biochemical compositions, including glucose, cholesterol, and total protein levels, were significantly changed in the treatment groups. Similarly, the ionic compositions of seminal fluid (Na, K, Ca, and Mg) also varied in the treatment groups. Furthermore, the 11-ketotestosterone levels were decreased, and the 17-ß estradiol levels were increased in the DEHP-treated groups. The mRNA expression levels of reproduction-related genes, including Fshr, Lhr, Ar, Erα, and Erß, were significantly changed in the DEHP-treated males in a dose-dependent manner. In conclusion, the findings of this study confirmed that environmentally relevant exposure to DEHP may contribute to a decline in the gamete quality of male fishes.
ABSTRACT
Aeromonas hydrophila is a conditional pathogen impacting public hygiene and safety. Hemolysin is a virulence factor of Aeromonas hydrophila that causes erythrocyte hemolysis, yet its transcriptional response to Cyprinus rubrofuscus remains unknown. Our investigation confirmed the hemolysis of hemolysin from A. hydrophila. Serum enzyme activity was evaluated weekly after C. rubrofuscus were immunized with hemolysin Ahh1. The results showed that the hemolysin enhances the serum superoxide dismutase (SOD), lysozyme (LZM), and catalase (CAT) activity, which reached a maximum on day 14. To elucidate the molecular interaction between hemolysin from A. hydrophila and the host, we performed transcriptome sequencing on the spleen of C. rubrofuscus 14 days post hemolysin infection. The total number of clean reads was 41.37 Gb, resulting in 79,832 unigenes with an N50 length of 1863 bp. There were 1982 significantly differentially expressed genes (DEGs), including 1083 upregulated genes and 899 downregulated genes. Transcript levels of the genes, such as LA6BL, CD2, and NLRC5, were significantly downregulated, while those of IL11, IL1R2, and IL8 were dramatically upregulated. The DEGs were mainly enriched in the immune disease, viral protein interaction with cytokine and cytokine receptor, and toll-like receptor pathways, suggesting that hemolysin stimulation can activate the transcriptional responses. RT-qPCR experiments results of seven genes, IL-8, STAT2, CTSK, PRF1, CXCL9, TLR5, and SACS, showed that their expression was highly concordant with RNA-seq data. We clarified for the first time the key genes and signaling pathways response to hemolysin from A. hydrophila, which offers strategies for treating and preventing diseases.
Subject(s)
Carps , Spleen , Animals , Aeromonas hydrophila , Hemolysin Proteins/genetics , HemolysisABSTRACT
A. hydrophila (Aeromonas hydrophila) is one of the most hazardous pathogenic microorganisms threatening the aquaculture industry and exhibits zoonotic-like characteristics. This study was designed to investigate the differential gene expression and pathway enrichment in the spleen of koi carp (Cyprinus carpio koi) upon A. hydrophila infection. The Illumina NovaSeq 6000 sequencing platform was used to identify 252 DEGs (differentially expressed genes), including 112 upregulated genes and 140 downregulated genes, in the spleens of koi carp challenged with A. hydrophila compared to those in the spleens of koi carp treated with PBS (phosphate-buffered saline). DEGs were shown to be involved in 133 pathways by KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis. Numerous immunological disease-related pathways, such as the immune defense network for IgA production, Staphylococcus aureus infection, and antigen processing and presentation, were enriched in the DEGs. In addition, the expression levels of 10 randomly screened DEGs, including the inflammatory factor nlrp3 (NOD-like receptor family pyrin domain containing 3), cytokine il-8 (interleukin-8), c2 (complement c2), c3 (complement c3), and the lipid mediator cox1 (cyclooxygenase-1), were compared by qPCR. The results showed that six genes, including il-8, cox1, and nlrp3, were upregulated according to both RNA-seq and qPCR validation, while four, including c2 and c3, showed downregulated expression. This result verified a strong correlation between the RNA-seq and qPCR datasets at the expression level. Moreover, this study provided splenic transcriptome data for koi carp during A. hydrophila infection and provided theoretical support for future drug development.
Subject(s)
Carps , Spleen , Animals , Carps/genetics , Aeromonas hydrophila , Interleukin-8 , NLR Family, Pyrin Domain-Containing 3 Protein , Gene Expression ProfilingABSTRACT
The skin represents an indicator of an animal's health status. Causes of cutaneous diseases in fish most often trace back to biological agents. However, fish skin diseases can also arise from a complex interaction of infectious and non-infectious causes, making it more difficult to identify a specific aetiology. In the period between April and September of the years 2019-2022, four koi carp (Cyprinus carpio koi) from two European countries presented with multifocal, irregularly round, few mm to 1 cm, variably raised cutaneous reddened areas. The fish displayed good general condition. Cutaneous samples, investigated by microbiological and molecular methods and microscopy, did not indicate a primary pathogenic agent. Gross and histological findings of the cutaneous biopsies were consistent with a multifocal/reactive process centred on dermal vessels. The histological features were reminiscent of angiomatosis, a benign proliferative condition affecting the dermal vessels of mammals, including human patients. The clinical-pathological presentation and the dermatologic condition that affected the koi carp are discussed and compared with the veterinary and human literature.
Subject(s)
Angiomatosis , Carps , Fish Diseases , Humans , Animals , Skin , Europe , MammalsABSTRACT
6-pyruvoyl-tetrahydropterin synthase (PTPS) is the second key enzyme of the pteridine biosynthetic pathway and it plays vital roles in fish body color formation. In this study, Ccptps of koi carp (Cyprinus carpio L.) was cloned, identified and characterized. The full-length cDNA of Ccptps was 1140 bp and encodes for 139 amino acids. Multiple alignments revealed that the amino acids sequence of CcPTPS shared the highest identity to that of C. carpio, and Ccptps was clustered with cyprinid fishes in phylogenetic tree. Liver tissues of koi carp exhibited the highest expression of Ccptps, followed by muscle and skin tissues. During early developmental stages, the expression of Ccptps declined from 2 dph to 4 dph, and increased from 4 dph to 12 dph. The expressions of Ccptps in three color-related tissues (skin, scale and caudal fin) of whole red (WR) koi carp were significantly higher than that of whole while (WW) koi carp. Immunohistochemistry results of skin tissues showed that CcPTPS was mainly located in epidermis, stratum compactum of dermis and muscle layer, with the signal intensities in stratum compactum and muscle layer were stronger in WR koi carp compared to WW koi carp. Co-expressions of CcPTPS, CcSPR and CcXDH were detected in skin tissues of WW and WR koi carps, with CcPTPS exhibited stronger signal intensity compared to CcSPR and CcXDH. These findings imply that Ccptps is potentially involved in koi carp body color formation through the pteridine synthesis pathway.
Subject(s)
Carps , Fish Diseases , Animals , Carps/genetics , Phylogeny , DNA, Complementary , Amino Acids , PteridinesABSTRACT
Economic importance of common carp (Cyprinus carpio L.) increases every year. Viral diseases are major threat for carp aquaculture and cause significant economic losses. Koi herpesvirus (KHV) is one of the most serious carp diseases. Current study is focused on confirmation of possible differences in early immune response to KHV depending on level of resistance. Class I interferon signalling, complement cascade and cell-mediated cytotoxicity are hypothesized as major mechanisms of early innate immune response against KHV. Different breeds of common carp show distinct level of resistance to KHV. Two breeds of common carp with completely different susceptibility to KHV were chosen for current research: amur wild carp (AS) as highly resistant and koi carp (KOI) as very susceptible breed. KHV infection caused no mortalities, but the viral load in selected tissues increased during infection. Levels of expressions of chosen genes was examined using qRT-PCR and overall change in protein expression profiles was analysed by mass spectrometry. Significant differences in immune response between AS and KOI were detected mostly at the level of protein expression. Although cell-mediated cytotoxicity showed minimal influence during KHV infection, many immune response parameters related to class I interferon signalling pathway and complement cascade were increased earlier during KHV infection in AS comparing to KOI.
Subject(s)
Carps , Fish Diseases , Herpesviridae Infections , Herpesviridae , Animals , Carps/genetics , Herpesviridae/physiology , Immunity , InterferonsABSTRACT
Galectins are ß-galactoside sugar binding proteins which function as important pattern recognition receptors (PRRs) in innate immunity. Here, we identified a galectin-9 gene from koi carp (Cyprinus carpio), named kGal-9. The ORF of kGal-9 is 963 bp in length, which encodes a polypeptide of 320 amino acids without either signal peptide. The predicted molecular weight is 36.25 kDa, and the isoelectric point is 8.3. Multiple sequence alignment showed that the putative kGal-9 contains two carbohydrate recognition domains (CRD), which are conserved in Galectin-9s. The phylogenetic tree showed that kGal-9 clustered to Galectin-9s from other teleosts, and shared the highest identity of 87.5% with Qihe crucian (Carassius auratus). kGal-9 mRNA was abundant in head kidney, gills, and gut, but low in liver and muscle. Further, the expression level of kGal-9 in the head kidney and liver increased significantly after Aeromonas veronii (abbreviated A.v) infection. Unexpectedly, kGal-9 showed a remarkable downregulation in the spleen at various time points post A.v infection. Intramuscular injection of pckGal-9 not merely reduced the bacterial load of spleen tissue, but also improved the survival rate of koi carp post A.v challenge. Besides, administration of pckGal-9 stimulated the expression of several immuno-related genes including proinflammatory cytokines (IL-1ß, IL-6), anti-inflammatory cytokine (IL-10), complement components (C4, C9), with fluctuation in spleen and head kidney. Taken together, the obtained results suggest that kGal-9 occupies an important role in innate immunity and defense against bacterial infection in koi carp.
Subject(s)
Carps , Fish Diseases , Aeromonas veronii/genetics , Amino Acids/genetics , Animals , Carbohydrates , Carps/genetics , Carps/metabolism , Fish Proteins/chemistry , Galectins/genetics , Galectins/metabolism , Immunity, Innate/genetics , Interleukin-10/genetics , Interleukin-6/genetics , Phylogeny , Protein Sorting Signals/genetics , RNA, Messenger/metabolism , SugarsABSTRACT
This study reports the occurrence of cyprinid herpesvirus 3 (CyHV-3) in koi carp (Cyprinus carpio koi) for the first time in India. The koi carp, with clinical signs of ulcer with haemorrhage on body surface, necrosis of fin and discolouration of gill associated with huge mortality, were observed in aquarium shops, rearing tanks and grow-out ponds located in Chennai, India. The PCR assay carried out on infected fish samples using different primer sets specific to CyHV-3 confirmed its presence in the infected fish. Sequence analysis of partial thymidine kinase gene revealed 100% similarity with the sequence of CyHV-3 available in GenBank. Cell lines of koi carp and catla were found to be susceptible to CyHV-3 and its replication was confirmed by viral-specific cytopathic effect, PCR and bioassay. The CyHV-3 infection was reproduced by intramuscular injection of inoculum prepared from CyHV-3-infected fish to satisfy Koch's postulates. Tissue tropism of CyHV-3 in infected fish by PCR assay revealed the presence of CyHV-3 in all vital organs with prominent band in gill and gut tissue.
Subject(s)
Carps , Fish Diseases , Herpesviridae Infections , Herpesviridae , Animals , Herpesviridae/genetics , Herpesviridae Infections/epidemiology , Herpesviridae Infections/metabolism , Herpesviridae Infections/veterinary , India/epidemiologyABSTRACT
Introduction: Koi carp, an ornamental fish derived from the common carp Cyprinus carpio (CC), is characterized by beautiful skin color patterns. However, the mechanism that gives rise to the characteristic vivid skin coloration of koi carp has not been clarified. The skin coloration of many teleosts changes in response to differences in the background color. This change in skin coloration is caused by diffusion or aggregation of pigment granules in chromatophores and is regulated mainly by sympathetic nerves and hormones. We hypothesized that there would be some abnormality in the mechanism of skin color regulation in koi carp, which impairs skin color fading in response to background color. Methods: We compared the function of melanin-concentrating hormone (MCH), noradrenaline, and adrenaline in CC and Taisho-Sanshoku (TS), a variety of tri-colored koi. Results and Discussion: In CC acclimated to a white background, the skin color became paler and pigment granules aggregated in melanophores in the scales compared to that in black-acclimated CC. There were no clear differences in skin color or pigment granule aggregation in white- or black-acclimated TS. The expression of mch1 mRNA in the brain was higher in the white-acclimated CC than that in the black-acclimated CC. However, the expression of mch1 mRNA in the brain in the TS did not change in response to the background color. Additionally, plasma MCH levels did not differ between white- and black-acclimated fish in either CC or TS. In vitro experiments showed that noradrenaline induced pigment aggregation in scale melanophores in both CC and TS, whereas adrenaline induced pigment aggregation in the CC but not in the TS. In vitro administration of MCH induced pigment granule aggregation in the CC but not in the TS. However, intraperitoneal injection of MCH resulted in pigment granule aggregation in both CC and TS. Collectively, these results suggest that the weak sensitivity of scale melanophores to MCH and adrenaline might be responsible for the lack of skin color change in response to background color in the TS.
Subject(s)
Carps , Epinephrine , Animals , Epinephrine/pharmacology , Melanophores/metabolism , Norepinephrine/pharmacology , Norepinephrine/metabolism , RNA, Messenger/metabolismABSTRACT
Microplastics (MPs) can be easily taken up by a wide range of aquatic animals and cause blockage of the digestive tract leading to starvation. Meanwhile, aquatic organisms are facing threats posed by food restriction in both wild and cultured environment. Little knowledge, however, exists on how MPs interact with food conditions to affect aquatic animals. Here, koi carp were exposed to polystyrene MPs (0, 100 or 1000⯵g/L) under controlled feeding (satiated or starved) for 30 or 60 days. MPs reduced and interacted synergistically with food conditions on growth after 30 days but antagonistically after 60 days. MPs reduced crude lipid and carbohydrate but increased and antagonistically interacted with feeding conditions on crude protein. Food conditions interacted with MPs on C, N and P but stoichiometric responses were decoupled with macromolecules changes. Food conditions antagonistically interacted with MPs on δ13C after 60 days. Linear discriminant analysis revealed that C:P and N:P were the two most important measured parameters accounting for the response of koi towards MPs and food restriction, presenting an antagonistic interaction of MPs and food status with the prolonged exposure duration.
Subject(s)
Carps , Water Pollutants, Chemical , Animals , Aquatic Organisms , Microplastics , Plastics , Water Pollutants, Chemical/analysis , Water Pollutants, Chemical/toxicityABSTRACT
Reports on abdominal tumours in koi carp are scarce and most are from the gonads. Their histological diagnosis is challenging due to the occurrence of mixed populations of neoplastic cells and the few availability of cross-reactive antibodies in fish tissues. The present study aims to provide a histopathological characterization of seventeen gonadal tumours, enriched by a wide antibody panel (vimentin, CD117, placental alkaline phosphatase-PLAP, AE1/AE3 cytokeratin, E-cadherin, proliferating cell nuclear antigen-PCNA, müllerian-inhibiting substance-MIS, GATA4 and Inhibin-α) applied on whole and tissue microarray (TMA) sections. Abdominal enlargement was associated with tumours filling 30%-80% of the abdominal cavity; frequently, the gonads had been completely replaced by neoplastic tissue. Twelve cases were characterized as sex cord-stromal tumours (SCSTs), three as germ cell tumours (GCTs), one as mixed germ cell sex cord-stromal tumour (MGCSCST) and one as carcinoma. By immunohistochemistry, PLAP enabled confirmation of GCTs, ovarian carcinoma and the objective identification of a further cell component in 8 out of the 12 SCSTs that were reclassified as mixed tumours. The use of an immunohistochemical panel can help in refining the histological diagnosis, but the morphological diagnosis still represents the main tool for the characterization of these tumours in koi carp.
Subject(s)
Carps , Fish Diseases/diagnosis , Neoplasms, Gonadal Tissue/veterinary , Animals , Female , Fish Diseases/pathology , Immunohistochemistry , Male , Neoplasms, Gonadal Tissue/diagnosis , Neoplasms, Gonadal Tissue/pathologyABSTRACT
Swim bladder disorders and consequent buoyancy problems are encountered in ornamental fish, including koi carp. Nevertheless, beyond clinical and pharmacological management, they are largely underdiagnosed. In this study, nine koi carp showing abdominal swelling and abnormal swimming behavior were investigated. Clinical approach, varying from case to case, included ultrasonographic and X-ray investigations, bacteriological analysis of the collected fluid, antimicrobial susceptibility pattern, and possibly histological analysis. Diagnostic imaging, corroborating gross examination, documented swim bladder deformation/dislocation and serous fluid within the swim bladder chambers of most animals. Bacteria belonging to the Aeromonas hydrophila/caviae group and Shewanella xiamenensis were identified. S. xiamenensis strains showed a sensibility to all tested molecules except for one strain, which was resistant to tetracycline and cyprofloxacin. Antibiotic treatment succeeded in the full recovery of three cases in which S. xiamemensis infection was detected. Chronic aerocystitis was histologically documented where tissue was available. The swim bladder histopathological findings highlighted a chronic process that had compromised the quality of life of the animals. A multidisciplinary clinical-pathological and microbiological approach is highly suggested to recognize swim bladder conditions as early as possible, aiming to drive medical intervention and raising the chances of fish survival.
ABSTRACT
MicroRNAs (miRNAs) are â¼22 nucleotide non-coding RNA molecules that act as crucial roles in plenty of biological processes. However, the molecular and cellular mechanisms of miRNAs to regulate skin color differentiation and pigmentation in fish have not been fully understood. Herein, we revealed that miR-206, a skin-enriched miRNA, regulates melanocortin 1 receptor (Mc1r, a key regulator of melanogenesis) expression by binding to its 3'-untranslated (UTR) region through bioinformatics and luciferase reporter assay in koi carp (Cyprinus carpio L.). The analysis of spatial and temporal expression patterns suggested that miR-206 is a potential regulator in the skin pigmentation process. Then, we silenced it in vivo with an antagomir method. The result showed a substantial increase of Mc1r mRNA expression and protein level, and also its downstream genes: tyrosinase (Tyr) and dopachrome tautomerase (Dct) that encoding key enzymes involved in melanin synthesis. Moreover, we constructed the miRNA-206 sponge lentivirus vector to transfect koi carp melanocytes in vitro, further checked the functions of melanocytes using Cck-8 and Transwell assays. As a result, inhibition of miR-206 significantly up-regulated Mc1r mRNA expression and protein level and accelerated the melanocyte proliferation and migration ability compared with the scrambled-sequence negative control group (miR-NC). Overall, these findings provide the evidence that miR-206 plays a regulatory role in the skin color pigmentation through targeting the Mc1r gene and would facilitate understanding the molecular regulatory mechanisms underlying miRNA-mediated skin color pigmentation in koi carp.
ABSTRACT
The indiscriminate use of antimicrobials in aquaculture results in antibiotic selection pressure and proliferation of antimicrobial resistant (AMR) bacteria. Frequent assessment of antimicrobial resistance in aquaculture environment is inevitable so as to reduce the passage of clinically important AMR from aquatic to other environment. The present study analysed the antimicrobial resistance of pathogens associated with diseased koi carp and goldfish from an ornamental fish farm. Phenotypic and genotypic characterization of the recovered isolates from both fishes revealed significant pathogens in aquaculture such as Aeromonas, Edwardsiella tarda, Acinetobacter, Lactococcus, Citrobacter, Enterobacter and Comamonas. Shannon-Wiener diversity of koi isolates (2·359) was found to be higher than that of goldfish (1·864). Antibiotic susceptibility testing using disc diffusion with 47 antibiotics revealed significant resistance pattern of Acinetobacter, Comamonas, Klebsiella and Enterobacter from goldfish and Edwardsiella, Aeromonas, Lactococcus, Enterobacter and Acinetobacter from koi with higher multiple antibiotic resistance indexes (>0·3). The minimum inhibitory concentration of antibiotics for the major resistant isolates was found to be very high with >256 µg. All the isolates were susceptible to amoxicillin, kanamycin, cefepime, cefexime, cefotaxime, ceftazidime, doripenem, ciprofloxacin and norfloxacin, recommending their successful application in the farm. SIGNIFICANCE AND IMPACT OF THE STUDY: Antimicrobial resistance is a major threat faced in aquaculture industry. The current study provides baseline information regarding the antibiotic resistance patterns of diverse pathogens recovered from ornamental koi carp and goldfish. The higher MAR index of pathogens and greater MIC of antibiotics for the resistant isolates highlighted the intense use of antibiotics in aquaculture farm. The potential of the pathogens to exhibit resistance even towards the new generation antibiotics remind the need of prudent use of antibiotics and continuous monitoring and surveillance programmes.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteria/genetics , Carps/microbiology , Fresh Water/microbiology , Goldfish/microbiology , Animals , Aquaculture , Bacteria/classification , Bacteria/drug effects , Bacteria/isolation & purification , Farms , Fishes/microbiology , Humans , Microbial Sensitivity TestsABSTRACT
This study investigated the impacts of various levels (0, 5, 10, 15, 20, and 25 g kg-1) of dietary ferula (Ferula asafoetida) powder (FP) on the hemato-immunological indices, antibacterial properties of skin mucus, survival rate, and growth performance as well as digestive enzyme activity of Koi carp, Cyprinus carpio koi, fingerlings. Following 63 days of feeding trail, WBCs, RBCs, Hb, hematocrit, MCV, MCH, MCHC, and lymphocyte levels increased coincident with an increasing FP level in experimental diets compared with control diet (P < 0.05). Dietary FP significantly increased total protein content and the activities of total immunoglobulins, lysozyme, and alternative hemolytic complement in a concentration-dependent manner in the serum of koi fish (P < 0.05). The elevation of the FP level in experimental diets resulted in an increase in SGR value and a reduction in FCR value (P < 0.05). The survival rate also increased significantly coincident with the increasing dietary FP level (P < 0.05). The results revealed that dietary ferula powder especially at levels of 20 and 25 g kg-1 could be used as an effective herbal dietary supplement in the enhancement of humoral innate immune responses and growth of koi carp.