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OBJECTIVE To establish an echocardiographic technique in koi carp (Cyprinus carpio), compare cardiopulmonary parameters under manual restraint versus anesthesia, and provide a gross anatomical and histologic cardiac description. METHODS A randomized, crossover echocardiography study was performed in 40 clinically healthy adult, unknown sex, privately owned koi carp on May 10 and 11 through June 26 and 27, 2021. Echocardiography was examined for each koi under manual restraint and isoeugenol at 50 ppm, with 3 measurements per examination performed by a radiologist and cardiologist. Two koi were euthanized for gross anatomic and histologic cardiac evaluation. RESULTS Mean ejection fraction (EF), stroke volume (SV), and cardiac output (CO) were significantly lower, mean heart rate (HR) was significantly higher, and opercular rate (OPR) was decreased significantly in anesthetized compared to manually restrained koi. Poor reproducibility for EF and SV was observed. CONCLUSIONS Echocardiography was feasible in both manually restrained and anesthetized koi; however, this technique may best be applied to monitoring trends over time in individual fish due to low reproducibility. Significant differences in multiple cardiopulmonary parameters, including HR, EF, SV, CO, and OPR, were present between manually restrained and anesthetized koi. A gross anatomic and histologic cardiac description is provided for this species to pair with the echocardiographic images. CLINICAL RELEVANCE This study provides the first description of echocardiography, cardiac gross anatomy, and histology in koi. The results support echocardiography as a safe and practical noninvasive diagnostic for cardiac assessment in koi under both manual restraint and anesthesia. [ABSTRACT FROM AUTHOR]

Animal skin patterns are increasingly explained using the Turing pattern model proposed by Alan Turing. The Turing model, a self-organizing model, can produce spotted or striped patterns. However, several animal patterns exist that do not correspond to these patterns. For example, the body patterns of the ornamental carp Nishiki goi produced in Japan vary randomly among individuals. Therefore, predicting the pattern of offspring is difficult based on the parent fish. Such a randomly formed pattern could be explained using a majority voting model. This model is a type of cellular automaton model that counts the surrounding states and transitions to high-number states. Nevertheless, the utility of these two models in explaining fish patterns remains unclear. Interestingly, the patterns generated by these two models can be detected among very closely related species. It is difficult to think that completely different epidermal formation mechanisms are used among species of the same family. Therefore, there may be a basic model that can produce both patterns. Herein, the Turing pattern and majority voting method are represented using cellular automata, and the possibility of integrating these two methods is examined. This integrated model is equivalent to both models when the parameters are adjusted. Although this integrated model is extremely simple, it can produce more varied patterns than either one of the individual models. However, further research is warranted to determine whether this model is consistent with the mechanisms involved in the formation of animal fish patterns from a biological perspective. [ABSTRACT FROM AUTHOR]

Aquaponics is a sustainable food production technology combining the tripartite relationship between fish, plants, and microorganisms. This study discusses the diversity and characteristics of bacteria communities in different aquaponic system compartments. Samples from red onion hydroponic compartment and koi biological filters were collected and analyzed for their bacterial community structure. Gene sequencing with high throughput 16S rRNA is used to analyze bacterial communities. Hydroponic compartments had significantly more bacterial communities than koi biological filters. The most dominant phylum in the hydroponic compartment is Proteobacteria (α, β, γ), Bacteroidota, Firmicutes, Planctomycetota, Cyanobacteria, and Actinobacteria. While in the biological filter are Bacteroidota, β-Proteobacteria, and Actinobacteria. The bacterial community in the hydroponic compartment showed significant specificity compared to the bacterial community in the biological filter. We also systematically describe bacterial communities in hydroponic compartments and biological filters and estimate their relevant functions in biological filtration of water. These results provide new insights into bacterial communities’ role in the hydroponic compartment for red onion growth as well as bacterial communities in the biofilter that act as bioremediation agents on water quality that affect the growth and survival of koi. However, it would be required a deeper analysis to identify its specific roles in aquaponics.

Introduction: Koi herpesvirus disease (KHVD) is attributed to cyprinid herpesvirus-3 (CyHV-3) and predominantly affects common carp and ornamental koi carp (Cyprinus carpio). This viral infection leads to substantial morbidity and mortality among these fish species. This study aimed to confirm the presence of KHVD in the Kurdistan region of Iraq by employing clinical and optimized molecular assays on fish populations experiencing high mortality among common carp in carp farms. Methodology: The present research was conducted in the Kalar district, situated at the heart of Garmian province in Iraqi Kurdistan. four samples from common carp fish farms were received by our laboratory. These samples specifically displaying clinical signs associated with koi herpesvirus (KHV) infection, were subjected to clinical examinations, and PCR assay in addition to sequence analysis. Results: The results of the current study revealed that the observed clinical signs, particularly gill necrosis, skin lesions, and sunken eyes, closely resembled the clinical signs of KHVD in common carp fish. In addition, PCR, nested PCR, and sequence analysis assay detected appropriate DNA fragments of the CyHV-3 major capsid protein gene confirming the first detection of KHVD in common carp fish in the Kurdistan region of Iraq. Conclusion: In this study, the results confirm the detection of KHVD in the Kurdistan region, Iraq, for the first time. This study revealed that CyHV-3 was responsible for KHVD-related signs and symptoms. Based on these results, it is strongly recommended that comprehensive studies be initiated to investigate the prevalence and distribution of CyHV-3. [ABSTRACT FROM AUTHOR]

Introduction: Carp edema virus (CEV) is a fish poxvirus that primarily infects the gills of common carp. CEV causes koi sleepy disease (KSD), which is highly contagious and can result in mortality of up to 100%. Methods: In the present study, we analyzed the stress and immune responses during KSD in two strains of common carp with different resistance to CEV: susceptible koi and resistant Amur sazan. Experiments were performed at two temperatures: 12°C and 18°C. In the case of koi carp, we also analyzed the effect of supplementation of 0.6% NaCl into tank water, which prevents mortality of the CEV-infected fish (salt rescue model). Results: We found that CEV-infected koi kept at 18°C had the highest viral load, which correlated with the most severe histopathological changes in the gills. CEV infection resulted in the activation of stress response reflected by the upregulated expression of genes involved in stress response in the stress axis organs and increased levels of cortisol and glucose in the blood plasma. These changes were the most pronounced in CEV-infected koi kept at 18°C. At both temperatures, the activation of antiviral immune response was observed in koi kept under freshwater and NaCl conditions upon CEV infection. Interestingly, a clear downregulation of the expression of adaptive immune genes was observed in CEV-infected koi kept under freshwater at 18°C. Conclusion: CEV induces a stress response and modulates adaptive immune response in koi, and this is correlated with the level of viral load and disease development. [ABSTRACT FROM AUTHOR]

Koi herpesvirus (KHV, CyHV-3) causes severe economic losses in carp farms. Its eradication is challenging due to the establishment of latency in blood leukocytes and other tissues. To understand the molecular mechanisms leading to KHV infection in leukocytes, common carp were bath-exposed to KHV at 17 °C. After confirming the presence of viral transcripts in blood leukocytes at ten days post infection, RNA-Seq was performed on peripheral blood leukocytes on the Illumina NovaSeq. KHV infection triggered a robust immune response mediated by pattern recognition receptors, mainly toll-like receptors (tlr2, tlr5, tlr7, and tlr13), urokinase plasminogen activator surface receptor-like, galectin proteins, and lipid mediators such as leukotriene B4 receptor 1. Enriched pathways showed increased mitochondria oxidative phosphorylation and the activation of signalling pathways such as mitogen-activated protein kinases (MAPKs) and vascular endothelial growth factor (VEGF). KHV-infected leukocytes showed low production of reactive oxygen species (ROS) and glutathione metabolism, high iron export and phagocytosis activity, and low autophagy. Macrophage polarization was deduced from the up-regulation of genes such as arginase non-hepatic 1-like, macrophage mannose receptor-1, crem, il-10, and il-13 receptors, while markers for cytotoxic T cells were observed to be down-regulated. Further work is required to characterise these leukocyte subsets and the molecular events leading to KHV latency in blood leukocytes. [ABSTRACT FROM AUTHOR]

Herpesviruses are common agents in animals of the aquatic environment. They infect many species of fish but only lead to disease in one or two species. Nevertheless, infected fish without clinical symptoms can actively transfer infectious agents to disease-susceptible species. The aim of the study was to identify and prove the natural presence of different herpesviruses. Koi, Nile tilapia, grass carp, goldfish and crucian carp were infected with a herpesvirus isolate 99% identical to goldfish herpesvirus (GHV) or cyprinid herpesvirus 2 (CyHV-2) obtained from crucian carp. Before and after infection, samples were collected non-lethally at different time points from all five fish species to identify and evaluate the replication of viruses naturally infecting the fish as well as the CyHV-2 experimentally infecting them. Gill swabs and separated leukocytes were subjected to PCR and the results compared. These samples yielded DNA of koi herpesvirus (KHV, also referred to as CyHV-3), GHV and a new herpesvirus. While Asian-lineage CyHV-3 DNA was detected in samples from crucian carp and goldfish, CyHV-2 DNA was found in samples from koi and tilapia. A new, hitherto unknown herpesvirus was identified in samples from grass carp, and was confirmed by nested PCR and sequence analysis. The survival rates were 5% for grass carp, 30% for tilapia, 55% for crucian carp, 70% for koi and 100% for goldfish at 20 days post infection. Evolutionary analyses were conducted and five clusters were visible: CyHV-1 (carp pox virus), CyHV-2 with sequences from koi and tilapia, CyHV-3 with sequences from crucian carp and goldfish, probable CyHV-4 from sichel and a newly discovered herpesvirus – CyHV-5 – from grass carp. The results obtained with the molecular tools as well as from the animal experiment demonstrated the pluripotency of aquatic herpesviruses to infect different fish species with and without visible clinical signs or mortality. [ABSTRACT FROM AUTHOR]

Insect meal has shown promise as a potentially sustainable source of nutrients for aquafeeds, offering an alternative to expensive and ecologically undesirable ingredients, in the context of population explosion and climate change. Despite this promising outlook, its effects on fish growth and immune responses remain to be thoroughly investigated. Our scientific goal was to experimentally test responses to replacements of the fish meal with a protein source derived from black soldier fly larvae meal (BSFLM). Possible impacts on growth, immunological response, and the expression of selected immune-system related genes were evaluated in Koi carp (Cyprinus carpio var. koi) using a biofloc culture system. Three hundred fish (20.0 ± 0.2 g) were allocated into five groups: a control group receiving a basal diet containing 0 g kg−1 BSFLM and four experimental groups in which fish meal was replaced with 50, 100, 150, and 200 g kg−1 BSFLM for eight weeks. After 4 weeks of feeding, there were no statistically significant differences in specific growth rate (SGR), feed conversion ratio (FCR), and survival rate between fish fed BSFLM-enriched diets at 50, 100, 150 g kg−1 and a control (0 g kg−1 BSFLM) diet. However, fish fed 200 g kg−1 BSFLM showed significantly improved weight gain (WG) and SGR compared to the control after 4 weeks; this difference persisted through 8 weeks (p < 0.05). After eight weeks, there was a moderate to weak negative linear regression shown in FCR (r = 0.470) and SR (r = 0.384), respectively, with the BSFLM levels, but significant and highly correlated linear relationships were observed in WG (r = 0.917) and SGR (r = 0.912). Immunological response analysis showed slight changes in lysozyme and peroxidase levels by replacing fish meal with BSFLM, but these apparent differences were not significantly related to experimental diets. Interestingly, mRNA transcripts of immune-related genes (TNF-α, TGF-β, IL1, IL10, and hsp70) were upregulated in the groups receiving higher amounts of BSFLM, with statistically significant differences observed in certain comparisons. Our findings reveal that fish meal can be effectively replaced by BSFLM, and that this not only has a positive effect on immune-related gene expression in Koi carp, but also on growth rate, pointing to the future potential role of BSFLM as an alternative fish meal protein in aquafeed formulation. [ABSTRACT FROM AUTHOR]

The number of exotic freshwater ornamental fish species released from the aquarium trade in Iran has been increasing in recent years. The Koi fish has already been reported from Iran, where it occurred in the southern Caspian Sea and Namak Lake basins. This is the first documented record of Cyprinus rubrofuscus from the Hormuz basin in southern Iran, which shows the range extension of this species in Iran. It seems there is no natural predator in most of the Iranian aquatic ecosystems for large Koi specimens. Therefore, the establishment of established populations in the natural lentic habitats is probable. Hence, eradication programs need to be accompanied by a public awareness campaign to ensure that the aquarium trade and hobbyists do not release these pet fishes into natural habitats. [ABSTRACT FROM AUTHOR]

This study aims to immunize the koi fingerlings immersion using the formalin-killed and freeze-dried E. coli DH5α carrying plasmid for the KHV DNA vaccine. 200 fish on each tank in a total water volume of 20 L. Each tanks consists of different vaccination group: PBS as control (10 mL; C), empty E. coli DHα (10 mL at 108 CFU mL-1; E), formalinkilled E. coli DHα:ORF81 (10 mL at 108 CFU mL-1; KE), freezedried E.coli DHα:ORF81 without formalin inactivation (10 mL at 108 CFU mL-1; FE), and formalin-killed and then freeze-dried E. coli DHα:ORF81 (10 mL at 108 CFU mL-1; KFE). The bath vaccination was conducted for 1 × 30 min. For the challenged test, fish were immersed with the 100 mL of LD50 dose of KHV (10–2 dilution from the KHV stock) for 30 min. The vaccination using E. coli DH5α:ORF81 could significantly modulate the innate and adaptive immunity of the fish and result in higher fish survival after KHV infection. The vaccination using formalin-killed or formalin-killed and freezedried E. coli DH5α:ORF81 will be further developed as an alternative to the costefficient koi or carp fingerlings vaccination method.

Lao Pa Koi (LPK) chicken is a popular fighting breed in Thailand, prized for (its unique characteristics acquired by selective breeding), and a valuable model for exploring the genetic diversity and admixture of red junglefowls and domestic chickens. In this study, genetic structure and diversity of LPK chicken were assessed using 28 microsatellite markers and mitochondrial DNA (mtDNA) D-loop sequences, and the findings were compared to a gene pool library from "The Siam Chicken Bioresource Project". High genetic variability was observed in LPK chickens using mtDNA D-loop haplotype analysis, and six haplotypes were identified. Microsatellite data revealed 182 alleles, with an average of 6.5 alleles per locus. These results confirmed the occurrence of genetic admixture of red junglefowl and Thai domestic chickens in LPK chicken breed. A maximum entropy modeling approach was used to analyze the spatial suitability and to assess the adaptive evolution of LPK chickens in diverse local environments. The model identified 82.52% of the area studied as unsuitable, and 9.34%, 7.11%, and 2.02% of the area indicated moderate, low, and high suitability, respectively. The highest contribution rate to land suitability for LPK chickens was found at an elevation of 100–250 m, suggesting the importance of elevation for their potential distribution. The results of this study provide valuable insights into the genetic origin of LPK chicken breed and identify resources for future genetic improvement. [ABSTRACT FROM AUTHOR]

The disease caused by carp edema virus (CEV) manifests with lethargy as a primary sign; this observation in koi in Japan gained the disease the name koi sleepy disease (KSD). In the years following the discovery of the virus in Japan, KSD cases have been noted in the UK in koi and common carp. Conducting research in order to expand knowledge of the processes of distribution of CEV in infected fish organs will be helpful for eradication and diagnostic purposes. Carp edema virus–affected fish with clinical signs of KSD were experimentally cohabited with common carp fry (30 fish). Three fish were euthanised by bath in a 0.5 g L−1 tricaine solution at one week intervals (7, 14, 21 and 28 days post cohabitation). Tissue samples from the brain, gills, spleen, kidney, intestines and skin were collected, and the total DNA was extracted and tested by real-time PCR. By the seventh day post infection, CEV DNA was most often found in the skin, gills and brain and less frequently in the kidney and intestines. In many of the common carp fry, CEV DNA could typically be found in several organs of each individual fish, although it was only found in one sample of spleen tissue. In this experimental study the pathogenesis of the CEV infection process was shown, the high infectivity of CEV was confirmed and the best organs were determined for sampling in CEV-infection experimentation. The real-time PCR method used in our cohabitation experiments was shown to be useful at the clinical and asymptomatic stage of virus infection. [ABSTRACT FROM AUTHOR]

Koi herpesvirus (KHV) and carp edema virus (CEV) are potential risks for the koi trade and for global common carp aquaculture. Due to the severe impact of both viruses, a robust diagnostic tool was required, capable of detecting and distinguishing both infections with high accuracy and precision. The aim of this study was to describe a realtime duplex TaqMan PCR assay for simultaneous detection of KHV and CEV in common carp and koi. Two pairs of primers with two TaqMan probes were used to amplify specific and conserved regions of KHV and CEV DNA. This assay was confirmed to be sensitive and specific. Limit of detections of the assay were 15 DNA copies/µL for KHV and 150 DNA copies/µL for CEV, respectively. This assay was able to identify and distinguish CEV and KHV, but was unable to identify other pathogens and sample matrices. For clinical validation, 18 KHV and CEV positive samples each, as well as 12 negative samples were tested with three different test methods, i.e., real-time duplex PCR, real-time simplex PCR, and conventional PCR. All three tests provide optimal conformity of results. The results showed that real-time duplex PCR was able to detect the presence and distinguish each pathogen in infected fish. This real-time duplex PCR assay is a rapid, sensitive, and specific test for detecting KHV and CEV in carp fish, thus it can be considered a valid alternative assay in aquaculture clinical laboratories. [ABSTRACT FROM AUTHOR]

Specimens of Gyrodactylus cyprini Diarova, 1964 (Monogenea: Gyrodactylidae) and Dactylogyrus extensus Mueller and Van Cleave, 1932 (Monogenea: Dactylogyridae) were obtained from the common carp Cyprinus carpio Linnaeus, 1758, which was reared in an outdoor pond at the Tamaki Field Station of the Fisheries Technology Institute, Mie Prefecture, Japan. Although G. cyprini is a well-known parasite of C. carpio in the Palearctic and USA and its existence in Japan was implied by the dermatitis examination of exported koi carp from Japan to Thailand, any specimen of G. cyprini has not been recorded in Japan. In addition, I determined some Japanese specimens previously identified as G. kherulensis Ergens, 1974 were also G. cyprini. This study obtained novel geographical records of these monogeneans, along with redescriptions of three monogenean species and molecular data of G. cyprini and D. extensus. [ABSTRACT FROM AUTHOR]

Carp edema virus disease (CEVD), also known as koi sleepy disease (KSD), represents a serious threat to the carp industry. The expression of immune-related genes to CEV infections could lead to the selection of crucial biomarkers of the development of the disease. The expression of a total of eleven immune-related genes encoding cytokines (IL-1β, IL-10, IL-6a, and TNF-α2), antiviral response (Mx2), cellular receptors (CD4, CD8b1, and GzmA), immunoglobulin (IgM), and genes encoding-mucins was monitored in gills of four differently KSD-susceptible strains of carp (Amur wild carp, Amur Sasan, AS; Ropsha scaly carp, Rop; Prerov scaly carp, PS; and koi) on days 6 and 11 post-infection. Carp strains were infected through two cohabitation infection trials with CEV genogroups I or IIa. The results showed that during the infection with both CEV genogroups, KSD-susceptible koi induced an innate immune response with significant up-regulation (p < 0:05) of IL-1β, IL-10, IL-6a, and TNF-α2 genes on both 6 and 11 days post-infection (dpi) compared to the fish sampled on day 0. Compared to koi, AS and Rop strains showed up-regulation of IL-6a and TNF-α2 but no other cytokine genes. During the infection with CEV genogroup IIa, Mx2 was significantly up-regulated in all strains and peaked on 6 dpi in AS, PS, and Rop. In koi, it remained high until 11 dpi. With genogroup I infection, Mx2 was up-expressed in koi on 6 dpi and in PS on both 6 and 11 dpi. No significant differences were noticed in selected mucin genes expression measured in gills of any carp strains exposed to both CEV genogroups. During both CEV genogroups infections, the expression levels of most of the genes for T cell response, including CD4, CD8b1, and GzmA were downregulated in AS and koi at all time points compared to day 0 control. The expression data for the above experimental trials suggest that both CEV genogroups infections in common carp strains lead to activation of the same expression pattern regardless of the fish's susceptibility towards the virus. The expression of the same genes in AS and koi responding to CEV genogroup IIa infection in mucosal tissues such as gill, gut, and skin showed the significant up-regulation of all the cytokine genes in gill and gut tissues from koi carp at 5 dpi. Significant down-regulation of CD4 and GzmA levels were only detected in koi gill on 5 dpi but not in other tissues. AS carp displayed significant up-expression of Mx2 gene in all mucosal tissues on 5 dpi, whereas in koi, it was upregulated in gill and gut only. In both carp strains, gill harbored a higher virus load on 5 dpi compared to the other tissues. The results showed that resistance to CEV could not be linked with the selected immune responses measured. The up-regulation of mRNA expression of most of the selected immune-related genes in koi gill and gut suggests that CEV induces a more systemic mucosal immune response not restricted to the target tissue of gills. [ABSTRACT FROM AUTHOR]

In this study, decoloration and detoxification of Astrazon Red FBL dye solutions using gamma rays have been examined at different environments (air, O2 saturated, 2.6 mM H2O2). In this context, the decoloration, mineralization, detoxification, pH, COD and BOD5 parameters were followed. Biodegradability ( 5 days of biological oxygen demand/ chemical oxygen demand -BOD5/COD-) ratio has been improved up on 2 kGy irradiation for all solutions. Optimum irradiation conditions were found to be 5 kGy pH 9 for air, 5 kGy pH 11 for O2 saturated, 7 kGy pH 11 for 2.6 mM H2O2 to provide mineralization and decoloration of Astrazon Red FBL solutions. Microtox bioluminescent test was conducted to measure the toxicity of Astrazon FBL solutions treated with high energetic rays. Toxicity reduction has been achieved at 5, 5 and 7 kGy in the range of 81.2, 86.7 and 56.3 % for air, O2 saturated and H2O2 solutions, respectively. As a result of study, it is found that Astrazon Red FBL dye could be treated by irradiation technology.

Background: Lymphatic neoplasia may occur in various types, such as lymphoma, lymphosarcoma, lympholeukemia, and plasmacytoid leukemia. Lymphoma, defined as a malignant tumour of lymphoid tissue, has been found in a number of fish families including Esocidae and Salmonidae. However, the occurrence of lymphoma is rare in those belonging to the Cyprinidae. A final diagnosis of ocular and testicular T-cell lymphoma in the present study was based on the clinical signs, morphology, and texture of the tumour masses in the macroscopic and microscopic examinations. In addition, histopathological and immunohistochemical findings corresponded to T-cell lymphoma characteristics. Case presentation: A 2-year-old hermaphrodite koi carp (Cyprinus carpio Linnaeus 1758) with a large ocular mass and severe exophthalmia in the right eye was referred to the Ornamental Fish Clinic in October 2020. Under anesthesia, enucleation was performed. 57 days after enucleation of the right eye, exophthalmia in the left eye was discovered. 221 days after surgery, the fish was discovered to be dead. At necropsy, a large soft tissue mass attached to the left testis was discovered. There were also small whitish nodules on the surface of the liver. Histopathology revealed a hypercellular ocular mass with scant connective tissue. The sections also revealed multifocal hemorrhages, round to ovoid neoplastic cells, mild-to-moderate anisokaryosis and anisocytosis, and mitotic figures. Basophilic neoplastic cells were found in blood vessels within the testicular mass, raising the possibility of systemic spread. The liver showed microscopic metastasis with morphologic similarities to the ocular and testicular tumors. The neoplastic cells infiltrating the left and right eyes as well as the testicular mass were immunohistochemically positive for CD3 but negative for CD20. The masses were diagnosed as T-cell lymphoma based on histopathological and immunohistochemical findings. Conclusions: This case report provides the first evidence of clinical, histopathological, morphological, and immunohistochemical findings of an ocular and testicular T-cell lymphoma in a hermaphrodite koi carp (Cyprinus carpio) in Iran. [ABSTRACT FROM AUTHOR]

Background: The accumulation of data on beneficial biological effects of probiotics and their metabolic products favors their potential use in the prevention and treatment of various malaises. Methods: Nine postmetabolites from Lactic acid bacteria (LAB) of human or dairy origin and their antiviral activity were studied using the cytopathic effect inhibition test. The virucidal capacity, their influence on the adsorption stage of Koi herpes virus (KHV) and their preventive role against subsequent viral challenge on intact Common carp brain (CCB) cells were also determined by titration assay. Residual viral infectivity in postmetabolites-treated samples was compared to mock-treated controls and Δlgs were calculated. Results: When administered during KHV replication, the microbial products isolated from Lactiplantibacillus plantarum showed remarkable activity with a selectivity index (SI) between 26.5 and 221.4, as those effects were dependent on the sample-virus incubation time. Postmetabolites from Lactobacillus gasseri and Lactiplantibacillus plantarum also demonstrated significant inhibition of KHV replication with SI of 24 and 16, respectively. The bioactive metabolites isolated from Limosilactobacillus fermentum had a minor effect on the viral replicative cycle. Compounds, produced during the fermentation by lactobacilli, grown on different nutritive media and collected at different time points, significantly inhibited extracellular KHV virions. All investigated postmetabolites remarkably blocked KHV attachment to the host cell (CCB), leading to a drop in viral titers by Δlg = 4.25–5.25, and exerted protective effects on CCB cells before they were subjected to viral infection. Conclusions: Our results open new horizons and promote LAB and their postbiotic products to be used in the prophylaxis and therapy of viral infections. [ABSTRACT FROM AUTHOR]

Common carp are among the oldest domesticated fish in the world. As such, there are many food and ornamental carp strains with abundant phenotypic variations due to natural and artificial selection. Hebao red carp (HB, Cyprinus carpio wuyuanensis), an indigenous strain in China, is renowned for its unique body morphology and reddish skin. To reveal the genetic basis underlying the distinct skin color of HB, we constructed an improved highfidelity (HiFi) HB genome with good contiguity, completeness, and correctness. Genome structure comparison was conducted between HB and a representative wild strain, Yellow River carp (YR, C. carpio haematopterus), to identify structural variants and genes under positive selection. Signatures of artificial selection during domestication were identified in HB and YR populations, while phenotype mapping was performed in a segregating population generated by HB×YR crosses. Body color in HB was associated with regions with fixed mutations. The simultaneous mutation and superposition of a pair of homologous genes (mitfa) in chromosomes A06 and B06 conferred the reddish color in domesticated HB. Transcriptome analysis of common carp with different alleles of the mitfa mutation confirmed that gene duplication can buffer the deleterious effects of mutation in allotetraploids. This study provides new insights into genotype-phenotype associations in allotetraploid species and lays a foundation for future breeding of common carp. [ABSTRACT FROM AUTHOR]

Koi sleepy disease (KSD) is a high mortality and infection viral disease caused by carp edema virus (CEV), which was a serious threat to aquaculture of common carp and export trade of Koi worldwide. Asymptomatic infection is an important cause of the difficulty in preventing KSD and its worldwide spread, because asymptomatic infection can be activated under appropriate condition. However, the understanding of the molecular correlates of these infections is still unknown. The purpose of this study was to compare the pathology change, enzyme activity, immunoglobulin activity, host and viral gene expression differences in acutely infected and cohabiting asymptomatic Koi infected with CEV. Healthy Koi were used as a control. The gross pathology, histopathology and ultrastructural pathology showed the difference and characteristics damage to the tissues of Koi under different infection conditions. Periodic Acid-Schiff stain (PAS), enzyme activity and immunoglobulin activity revealed changes in the immune response of gill tissue between acutely infected, asymptomatic infected and healthy Koi. A total of 111 and 2484 upregulated genes and 257 and 4940 downregulated genes were founded in healthy Koi vs asymptomatic infected Koi and healthy Koi vs acutely infected Koi, respectively. Additionally, 878 upregulated genes and 1089 downregulated genes were identified in asymptomatic vs. acutely infected Koi. Immune gene categories and their corresponding genes in different comparison groups were revealed. A total of 3, 59 and 28 immune-related genes were identified in the group of healthy Koi vs asymptomatic infected Koi, healthy Koi vs acutely infected Koi and asymptomatic infected Koi vs acutely infected Koi, respectively. Nineteen immune-related genes have the same expression manner both in healthy Koi vs acutely infected Koi and asymptomatic Koi vs acutely infected Koi, while 9 immunerelated genes were differentially expressed only in asymptomatic Koi vs acutely infected Koi, which may play a role in viral reactivation. In addition, 8 differentially expressed genes (DEGs) were validated by quantitative reverse transcription PCR (RT-qPCR), and the results were consistent with the RNA-Seq results. In conclusion, the data obtained in this study provide new evidence for further elucidating CEV-host interactions and the CEV infection mechanism and will facilitate the implementation of integrated strategies for controlling CEV infection and spread. [ABSTRACT FROM AUTHOR]

This paper showcases the kaupapa (philosophy) and practices of a Māta Waka (pan-tribal), community-based Kaupapa Māori service provider in the nation-state currently known as New Zealand. Te Hou Ora Whānau Services aims to provide services that support and empower tamariki (children), rangatahi (youth), and whānau (extended families) from diverse backgrounds and experiential realities to fulfil their potential within the context of their cultural heritage and their communities. The purpose of this study was to explore the philosophy and values underpinning the everyday practices and experiences of eleven kaimahi (practitioners) who work for the organisation. Analyses of the data identified five overarching pou, or foundational supports that underlie kaimahi ways of being, knowing, relating, and doing, together with seven 'takepū,' or preferred ways of engaging with others. Findings of the study provide an important window into the holistic, relational kaupapa of this Māta Waka organisation and its kaimahi: a set of commitments and actions that, as the findings demonstrate, are fundamentally a practice of aroha (love). While some of the elements are specific to the New Zealand context, the pou and takepū offer guidance relevant to programs globally seeking to successfully and creatively respond to the priorities, aspirations, and moemoeā/dreams of Indigenous children, young people, their families, and communities. [ABSTRACT FROM AUTHOR]

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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 blackacclimated 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. [ABSTRACT FROM AUTHOR]

Koi herpesvirus (KHV) is a highly contagious virus that causes high mortality in koi and common carp, leading to a reduction in production worldwide. Recent diagnostic tests based on molecular methods alone (nucleic acid amplification) and indirect immunoassay methods (antibody detection) can be confirmed over KHV infections or prior exposure and latent infections. Unfortunately, there is no established method to detect KHV virus particles, especially when virus titers are low. Therefore, we propose an alternative, direct immunoassay method for viral detection using a single-chain variable fragment (scFv), a specific region of IgG antibodies that binds specifically to KHV particles. The results of functional analyses indicated that four putative scFv candidates, C5, F8, F6, and E4, were specific to KHV, but only F6 and C5 had a high binding affinity. The binding characteristics were confirmed by indirect competitive and sandwich enzyme-linked immunosorbent assays, which indicated that F6 and C5 have a broad penetration area to the binding region and share a similar epitope with commercial KHV monoclonal antibodies. These characteristics were further confirmed by their interactions with purified KHV coat protein by indirect ELISA and Western blot analyses. In conclusion, the F6 and C5 scFvs have adequate binding affinity to KHV particles to permit their use in immunoassays. [ABSTRACT FROM AUTHOR]

Simple Summary: The diversity of fish resources is an important component of biodiversity. As a branch of fish, the diversity of koi varieties is conducive to improving the genetic quality of offspring, avoiding inbreeding, and improving their adaptability to the natural environment. The variety classification of koi is a necessary step to improve the diversity of koi varieties and breeding quality. The traditional manual classification method of koi variety faces some problems, such as high subjectivity, low efficiency, and high misclassification rate. Therefore, we studied an intelligent method of classifying koi variety using an artificial intelligence approach, and designed a deep learning network model, KRS-Net. The intelligent and nondestructive classification was realized for 13 varieties of koi by using the proposed model, and the accuracy rate was 97.9%, which is higher than that of the classical mainstream classification network. This study provides a reference for intelligent classification of marine organisms, and can be extended to the screening and breeding of other species. As the traditional manual classification method has some shortcomings, including high subjectivity, low efficiency, and high misclassification rate, we studied an approach for classifying koi varieties. The main contributions of this study are twofold: (1) a dataset was established for thirteen kinds of koi; (2) a classification problem with high similarity was designed for underwater animals, and a KRS-Net classification network was constructed based on deep learning, which could solve the problem of low accuracy for some varieties that are highly similar. The test experiment of KRS-Net was carried out on the established dataset, and the results were compared with those of five mainstream classification networks (AlexNet, VGG16, GoogLeNet, ResNet101, and DenseNet201). The experimental results showed that the classification test accuracy of KRS-Net reached 97.90% for koi, which is better than those of the comparison networks. The main advantages of the proposed approach include reduced number of parameters and improved accuracy. This study provides an effective approach for the intelligent classification of koi, and it has guiding significance for the classification of other organisms with high similarity among classes. The proposed approach can be applied to some other tasks, such as screening, breeding, and grade sorting. [ABSTRACT FROM AUTHOR]

OBJECTIVE To evaluate the effect of a multidose acyclovir protocol on koi herpesvirus (KHV) viral load and mortality in a cohabitation challenge. ANIMALS 180 koi fish. PROCEDURES Forty fish (shedders) were immersed in a 0.5 KHV plaque-forming units/mL static bath for 8 hours. Mock shedders were treated similarly but exposed to cell culture media. KHV shedders were then transferred into 8 tanks (5 shedders per tank) containing 10 naïve fish (cohabitants) each. Fish in the acyclovir group (AT) received a 10 mg/kg acyclovir intracoelomic injection 1, 3, and 6 days after the first confirmed KHV mortality. Positive controls (PC) were treated similarly but received sterile saline injections. Negative controls (NC) were exposed to mock shedders. Morbidity and mortality were evaluated daily for 50 days post-challenge. Quantitative PCR was used to determine viral load in the gill biopsies of shedders and cohabitants collected at days 19 (T1), 22 (T2), 25 (T3), 34 (T4), and 50 (T5) post-challenge. RESULTS Survival curves analyzed by the Gehan-Breslow-Wilcoxon method revealed a delayed onset of mortalities and a significantly lower KHV load at T2 and T3 detected in AT cohabitant fish (P = .042) compared to PC group. However, there were no significant differences in overall mortality or viral loads at T5. CLINICAL RELEVANCE The acyclovir protocol used in this study did not control viral infection or mortality at the end of the 50-day trial. Shorter intervals between injections could improve outcomes, but the additional stress inflicted by handling should be considered. Exploring other therapeutic alternatives and doses is warranted. [ABSTRACT FROM AUTHOR]

Plant polyphenol supplementation may improve fish health in aquaculture systems. To assess the potential benefits and function mechanism of plant polyphenols in aquaculture, fish were fed either basal feed (CON) or the basal feed supplemented with 500 mg/kg of curcumin (CUR), oligomeric proanthocyanidins (OPC), chlorogenic acid (CGA), or resveratrol (RES). After an 8-week feeding experiment, blood samples were used to analyze the concentrations of biochemical indices. Gut samples were collected to evaluate microbiota, short chain fatty acid (SCFA) levels, and gene expression. The results indicated that polyphenol administration reduced serum glucose and insulin. Lysozyme activity was enhanced by OPC and CGA, and superoxide dismutase activity was increased by CUR, OPC, and CGA. The gut microbial structure of the RES group was segregated from that of the CON, and the genus Bacteroides was identified as a potential biomarker in the CUR, CGA, and RES groups. Total gut SCFA increased in the CUR, CGA, and RES groups. A strong correlation was observed between Bacteroides and SCFA. In conclusion, dietary polyphenols have distinct anti-inflammatory, anti-oxidant, and anti-hyperglycemic activities that may be closely associated with their microbiota-modulation effects. [ABSTRACT FROM AUTHOR]

Since the end of the1990ies, Cyprinid herpesvirus 3 (also known as koi herpesvirus, KHV) has caused mass mortality events of koi and common carp all over the globe. This induced a high economic impact, since the KHV disease cannot be cured up to now, but only prevented by vaccination. Unfortunately, there is only one commercial vaccine available which is not approved in most countries. Therefore, there is an urgent need for new, safe and available vaccines. In this study, a live attenuated vaccine virus was generated by cell culture passages of virulent KHV, and shown to protect carp or koi after immersion or oral application against wild type challenge. An advantage of boost immunization was demonstrated, especially after oral application. Vaccination induced no or mild clinical signs and protecting antibodies have been measured. Additionally, the vaccine virus allowed differentiation of infected from vaccinated animals (DIVA) by PCR. The attenuation of the newly generated vaccine was tracked down to a partial deletion of open reading frame 150. This was confirmed by the generation of engineered ORF150 deletion mutants of wild-type KHV which exhibited a similar attenuation in vivo. [ABSTRACT FROM AUTHOR]

Quality evaluation of koi (Cyprinus rubrofuscus) is essential to the koi industry. However, the community has a lack of knowledge about the quality evaluation of koi and about features of koi which are valuable. During the "All Japan Nishikigoi Show" competition, a quality evaluation of koi was conducted by authoritative experts. In the present study, photos of participants of this competition were used for analysis to discover features affecting the quality of the Kohaku variety of koi. The HSVA color model was used to define and extract from image red and pale red coverage rates in Kohaku body coloration. Body aspect ratio, body proportions, red coverage rate, and pale red coverage rate were extracted using tools written in Python programming language. Analyses of the extracted data revealed the significant impact body aspect ratio (p < 0.001) and pale red coverage rate (p < 0.001) on Kohaku quality. A low body aspect ratio has a positive impact on Kohaku quality, while a high pale red coverage rate has a very strong negative impact on Kohaku quality. This study and its results can be an important foundation for further research about koi quality. [ABSTRACT FROM AUTHOR]

The koi carp is an ornamental fish that was obtained through artificial selection from the common carp (Cyprinus carpio L.). The most economically important traits of koi are their beautiful skin patterns in bright colors. As seasonality is an important factor in the biology and ecology of fish, we thus assumed that seasonal changes are involved in regulating the formation of skin color and patterns of koi carp. The white, red, cyan, and black skin colors from four varieties of scaleless koi carp (Doitsu Shiromuji (W), Doitsu Kohaku (W R), Doitsu Showa Sanke (WRI), and Kumonryu (WI)) were evaluated using the CIELab color space (lightness, redness, and yellowness) in different seasons. Compared to winter, the yellowness of the white color in all koi varieties decreased in summer and autumn. The black skin color areas in WRI and WI koi increased in summer and autumn compared to winter. The yellowness of the red color decreased only in WRI koi, while no changes were observed in W R koi. Targeted metabolomics analysis revealed that the levels of the structural pigment guanine of all koi varieties showed significant seasonal variation. Of seven detected carotenoids, the zeaxanthin and tunaxanthin contents in W, WI, and WRI koi changed with the seasons, while none of the carotenoids in W R koi were altered. Of the seven potential regulatory metabolites, epinephrine, melatonin, and cyclic adenosine monophosphate (cAMP) in all four koi varieties showed the highest levels in winter. A correlation analysis suggested that the seasonal changes in white skin color occurred through the epinephrine-cAMP pathway; melanin-dependent and carotenoid-dependent skin color changes occurred through melatonin in koi carp. This study demonstrated the seasonal plasticity of skin color in koi carp regulated by melatonin and epinephrine, associating with variety and color specificity. [Display omitted] • Koi variety differences in seasonal skin color plasticity. • Components of seasonal variation in carotenoids are zeaxanthin and tunaxanthin. • Seasonality changes the red skin of koi through melatonin. • Seasonality changes the white skin of koi through epinephrine-cAMP pathway. [ABSTRACT FROM AUTHOR]

Klebsiella michiganensis is emerging as an important human pathogen of concern especially strains with plasmid-mediated carbapenemase genes. The IncX3- bla NDM-5 plasmid has been described as the primary vector for bla NDM-5 dissemination. However, whether strains with this plasmid have any competitive edge remain largely unexplored. We characterized a bla NDM-5-producing Klebsiella michiganensis strain (KO_408) from Japan and sought to understand the driving force behind the recent dissemination of IncX3- bl aNDM-5 plasmids in different bacterial hosts. Antibiotic susceptibility testing, conjugation, and whole-genome sequencing were performed for KO_408, a clinical isolate recovered from a respiratory culture. Fitness, stability, and competitive assays were performed using the IncX3- bla NDM-5 plasmid, pKO_4-NDM-5. KO_408 was ascribed to a novel sequence type, ST256, and harbored resistance genes conforming to its MDR phenotype. The bla NDM-5 gene was localized on the ~44.9 kb IncX3 plasmid (pKO_4-NDM-5), which was transferable in the conjugal assay. The acquisition of pKO_4-NDM-5 did not impose any fitness burden and showed high stability in the host cells. However, transformants with pKO_4-NDM-5 were outcompeted by their host cells and transconjugants with the IncX3- bla OXA-181 plasmid. The genetic environment of bla NDM-5 in pKO_4-NDM-5 has been previously described. pKO_4-NDM-5 showed a close phylogenetic distance with seven similar plasmids from China. KO_408 clustered with strains within the KoI phylogroup, which is closely associated with carbapenemase genes. This study highlights the emergence of a high-risk Klebsiella michiganensis clone harboring carbapenemase genes and affirms that the recent spread of IncX3- bla NDM-5 plasmids might be due to their low fitness cost and stability but not their competitive prowess. [ABSTRACT FROM AUTHOR]

A ten-week feeding trial was performed to evaluate the effects of chlorogenic acid (CGA) on the growth, body color, antioxidant activity, serum biochemical parameters, and immune response of koi. Five diets were formulated to contain 0 (control), 200 (CGA-2), 400 (CGA-4), 600 (CGA-6), and 800 (CGA-8) mg/kg CGA for the rearing of experimental fish. By means of an analysis by polynomial orthogonal contrasts, it was determined that dietary supplementation with CGA had no significant effects on the growth performance of koi (P > 0.05), while body color indices (redness, yellowness, and lightness) showed a linear order polynomial (P < 0.05). A quadratic response was found for the relationship between total carotenoid in scale of koi and dietary CGA levels, with an optimum level of 381.84 mg/kg. Dietary CGA levels and lysozyme activity showed a cubic relationship polynomial model, with an optimum level of 383.89-975.73 mg/kg. TGF-β expression in the head-kidney of koi was obviously improved by 600 mg/kg CGA diet (P < 0.05). And in regards to effect in serum, SOD and CAT activities was best explicated by a linear order polynomial, while the quadratic and cubic relationship polynomial models represented the GSH and GSH-Px activities, which the optimal supplementation level of CGA was estimated at 550.78 and 231.74-782.66 mg/kg, respectively. The relationship between ALT activity and CGA supplementation was best explicated by quadratic relationship polynomial, with an optimum level of 522.78 mg/kg. Based on the model observed, 522.78-600 mg/kg CGA was the recommended supplementation in diet that improved the body color, antioxidant activity, biochemical parameters, and immune response of koi. [ABSTRACT FROM AUTHOR]

Background. Myxosporean diversity is a hot topic since they are difficult to accurately identify and classify. Many Myxobolus parasites have been named as Myxobolus koi because of their similar morphological features with the species originally reported. However, the distinctions in fine morphological features, host specificity, and molecular data have given rise to the attention of researchers. Methods. The classical morphometric and histological methods were used to describe the Myxobolus dajiangensis n. sp. in morphology. The common techniques in modern molecular biology and the methods of phylogenetic analyses were combined to identify the species. Results. Plasmodia of interlamellar-vascular type were found in the vascular network of gill lamellae. Mature myxospores of M. dajiangensis n. sp. were elongated and pyriform from the frontal view. The myxospores were 14.8 ± 0.4 (13.9-15.6) mm in length, 8.0 ± 0.5 (7.2-9.1) mm in width, and 5.5 mm in thickness. The two polar capsules were pyriform and slightly different in length. The length of the larger polar capsules was 8.0 ± 0.4 (7.1-8.8) mm, and it was 7.4 ± 0.4 (6.1-8.0) mm for the smaller ones. The width of both polar capsules was 2.5 ± 0.2 (2.0-3.2) mm. The polar filaments within the polar capsules were each coiled nine to 11 turns. Comparative analysis of both the morphological and molecular data between the present speices and other similar species revealed that the present species is a novel species, Myxobolus dajiangensis n. sp. Also, M. koi (FJ710800) was misidentified and the congener with M. dajiangensis n. sp., depending on the secondary structures of SSU rRNA and phylogenetic analysis. Moreover, the cryptic species existed in the M. koi parasites. [ABSTRACT FROM AUTHOR]

The increasing number of Koi ornamental fish interested around the world, this fish farming business is increasingly developing. Blitar is one of the geographically suitable areas to develop Koi fish farming. The resulting production has a quality of Koi better than from other countries. Cultivation models and techniques developed are also increasingly advanced, but often cultivators have disease problems stemming from decreasing environmental quality that are not handled early. This attracted the Brawijaya University Doctoral community service team to help find solutions to these problems.

Klebsiella michiganensis is emerging as an important human pathogen of concern especially strains with plasmid-mediated carbapenemase genes. The IncX3-blaNDM-5 plasmid has been described as the primary vector for blaNDM-5 dissemination. However, whether strains with this plasmid have any competitive edge remain largely unexplored. We characterized a blaNDM-5-producing Klebsiella michiganensis strain (KO_408) from Japan and sought to understand the driving force behind the recent dissemination of IncX3-blaNDM-5 plasmids in different bacterial hosts. Antibiotic susceptibility testing, conjugation, and whole-genome sequencing were performed for KO_408, a clinical isolate recovered from a respiratory culture. Fitness, stability, and competitive assays were performed using the IncX3-blaNDM-5 plasmid, pKO_4-NDM-5. KO_408 was ascribed to a novel sequence type, ST256, and harbored resistance genes conforming to its MDR phenotype. The blaNDM-5 gene was localized on the ~44.9 kb IncX3 plasmid (pKO_4-NDM-5), which was transferable in the conjugal assay. The acquisition of pKO_4-NDM-5 did not impose any fitness burden and showed high stability in the host cells. However, transformants with pKO_4-NDM-5 were outcompeted by their host cells and transconjugants with the IncX3-blaOXA-181 plasmid. The genetic environment of blaNDM-5 in pKO_4-NDM-5 has been previously described. pKO_4-NDM-5 showed a close phylogenetic distance with seven similar plasmids from China. KO_408 clustered with strains within the KoI phylogroup, which is closely associated with carbapenemase genes. This study highlights the emergence of a high-risk Klebsiella michiganensis clone harboring carbapenemase genes and affirms that the recent spread of IncX3-blaNDM-5 plasmids might be due to their low fitness cost and stability but not their competitive prowess.

The need for yield research is increasing with the popularity of aquaponic food production systems where plants and fish are grown together in a recirculating system. Our objective was to compare the yield of three basil cultivars ("Elenora," "Genovese," "Nufar," Ocimum basilicum) in four configurations (greenhouses: floating raft DWS, ebb & flow, A‐frame; warehouse: floating raft; 1–8 tank replications/treatment) as well as a soilless control to establish a baseline for aquaponic producers in northern latitudes. We also tested whether basil yield varied among fish types: yellow perch, Perca flavescens, goldfish, Carassius auratus, tilapia, Oreochromis spp., and koi, Cyprinus carpio. Fish tank water temperature, nitrite, and alkalinity levels differed significantly over seasons (summer, winter). There was a significant decrease in basil production in all aquaponic systems that used koi (greenhouse, warehouse). No significant difference in basil yield was found among greenhouse and aquaponic systems for perch, tilapia, or goldfish nor among the basil "Elenora," "Genovese," and "Nufar." Considerable variation occurred in yield although most of it occurred in fresh weights, being reduced significantly in dry weights. The effects of growing koi fish on reducing aquaponic production of basil is noteworthy, although future research is needed to the exact cause. [ABSTRACT FROM AUTHOR]

To maximize profits from aquaponic system, a high-price 'koi' carp (Cyprinus carpio) produced at Sultan Qaboos University was used. This study consisted of two experiments. The first experiment aimed to determine the optimal stocking density of 'koi' carp in an aquaponic system. In the second experiment, the optimal density of 'koi' carp based on the first experiment was used in combination with tomato, eggplant, and mint as a control from the first experiment. The aquaponic system consisted of 9 pairs of glass tanks; in each pair water was recirculating with the use of a submersible pump. In the first experiment, 'koi' carp fry (0.4-0.5 g each) were stocked at 50, 100 and 150 individuals per tank (0.1 m3) in combination with mint and the ornamental plant Petunia. Fish were stocked in tanks with and without plants. Results show that 'koi' carp stocked at 100 per tank without plants had significantly better growth (mean final weight of 7.5 g from the initial weight of 0.4-0.5 g). Fish stocked in tanks with plants, 'koi' carp at 50 per tank had significantly better growth (mean final fish weight of 5.2 g from the initial weight of 0.4-0.5 g). Survival rate was highest at 50 per tank with or without plants. Mint growth was better in tanks with 50 fish per tank. Petunia had flowers in all treatments. Water quality parameters were within optimal range even at high fish stocking densities. Overall, stocking density of 50 fish per tank can be recommended based on survival data. In the second experiment, 30 'koi' carp juveniles were stocked in all tanks without plants (i.e. 6.5 g, 5.5 g and 6.1 g respectively) for each treatment in combination with mint, tomato, and eggplant. Growth and survival rates of koi (30 fish per tank) in combination with mint, eggplant and tomato were high with a weight gain of 21.3, 17.6, 17.6 g and survival rate of 90%, 95.5% and 87.8%, respectively. The results showed that 'koi' can be successfully grown with different plants in an aquaponic system. [ABSTRACT FROM AUTHOR]

Gills of fish are involved in respiration, excretion and osmoregulation. Due to numerous interactions between these processes, branchial diseases have serious implications on fish health. Here, "koi sleepy disease" (KSD), caused by carp edema virus (CEV) infection was used to study physiological, immunological and metabolic consequences of a gill disease in fish. A metabolome analysis shows that the moderately hypoxic-tolerant carp can compensate the respiratory compromise related to this infection by various adaptations in their metabolism. Instead, the disease is accompanied by a massive disturbance of the osmotic balance with hyponatremia as low as 71.65 mmol L−1, and an accumulation of ammonia in circulatory blood causing a hyperammonemia as high as 1123.24 µmol L−1. At water conditions with increased ambient salt, the hydro-mineral balance and the ammonia excretion were restored. Importantly, both hyponatremia and hyperammonemia in KSD-affected carp can be linked to an immunosuppression leading to a four-fold drop in the number of white blood cells, and significant downregulation of cd4, tcr a2 and igm expression in gills, which can be evaded by increasing the ion concentration in water. This shows that the complex host-pathogen interactions within the gills can have immunosuppressive consequences, which have not previously been addressed in fish. Furthermore, it makes the CEV infection of carp a powerful model for studying interdependent pathological and immunological effects of a branchial disease in fish. [ABSTRACT FROM AUTHOR]

This research aims to study the ability of Aloe vera (Burm.f.), extract to treat Koi fry [Cyprinus rubrofuscus (Lacépède, 1803)] which is attacked by Aeromonas hydrophila (Chester, 1901). The method used was the experimental method using a completely randomized design with five treatments and three replications was applied. The treatment used was immersion with A. vera extract with a concentration of treatment A (0 mg kg-1), B (150 mg kg-1), C (300 mg kg-1), D (450 mg kg-1) and E (600 mg kg-1). The variables observed were in vitro test, LC50-24-h test, clinical symptoms which included damage to fish body, fish feeding response and fish response to shock which were analyzed descriptively comparatively. The results showed A. vera extract has the ability as an antibacterial with inhibition zone diameter reaching 20.49 mm at a concentration of 5 000 mg kg-1 and LC50 test 48 h (in vivo test) of 1 157.16 mg kg-1, so for the treatment of fish use concentrations below that concentration. The use of aloe vera extract with a concentration of 300 mg kg-1 for 48 h was effective to treat koi fry infected with A. hydrophila by healing relatively fast body damage, response to feed and shock is normal and fast. [ABSTRACT FROM AUTHOR]

Koi herpesvirus (KHV) is highly contagious and lethal to cyprinid fish, causing significant economic losses to the carp aquaculture industry, particularly to koi carp breeders. Vaccines delivered through intramuscular needle injection or gene gun are not suitable for mass vaccination of carp. So, the development of cost-effective oral vaccines that are easily applicable at a farm level is highly desirable. In this study, we utilized chitosan-alginate capsules as an oral delivery system for a live probiotic (Lactobacillus rhamnosus) vaccine, pYG-KHV-ORF81/LR CIQ249, expressing KHV ORF81 protein. The tolerance of the encapsulated recombinant Lactobacillus to various digestive environments and the ability of the probiotic strain to colonize the intestine of carp was tested. The immunogenicity and the protective efficacy of the encapsulated probiotic vaccine was evaluated by determining IgM levels, lymphocyte proliferation, expression of immune-related genes, and viral challenge to vaccinated fish. It was clear that the chitosan-alginate capsules protected the probiotic vaccine effectively against extreme digestive environments, and a significant level (P < 0.01) of antigen-specific IgM with KHV-neutralizing activity was detected, which provided a protection rate of ca. 85% for koi carp against KHV challenge. The strategy of using chitosan-alginate capsules to deliver probiotic vaccines is easily applicable for mass oral vaccination of fish. [ABSTRACT FROM AUTHOR]

The skin color of koi carp (Cyprinus carpio L.) is one of the traits that most influence their ornamental and economic values. The present study suggested the effects of temperature fluctuation on koi carp in terms of skin color and plasma carotenoids and related-metabolites. The main results were as follows. (1) The vulnerability of koi skin color to acute temperature stress was in the order of white koi> black koi> yellow koi. Both high- (25°C-30°C-25°C) and low-temperature (25°C-20°C-25°C) fluctuations tended to decrease the saturation of white koi. The temperature fluctuation had little effects on the skin color of black and yellow koi. (2) Targeted metabolomics analysis indicated that the effects of cooling stress on oxycarotenoids of all five koi varieties were reversible. The plasma oxycarotenoids in mirror koi with all colors were insensitive to acute heat stress. However, the cooling process from a high temperature (30°C-25°C) still made contributions to the increase of oxycarotenoids. (3) The principal component analysis confirmed the deviation of carotenoid-related metabolites after high temperature fluctuation and the reversibility after low temperature fluctuation. Finally, the correlation analysis revealed that koi skin brightness was negatively correlated with the plasma guanine content and that temperature fluctuations might change koi skin brightness via the L(-)-epinephrine-guanine pathway. The red hue and yellow hue were negatively correlated with the oxycarotenoids in plasma, suggesting that oxycarotenoids were favorable for enhancing koi skin color saturation. Overall, this study revealed the direct action of temperature fluctuations on the skin color and carotenoid-related metabolites of koi. [Display omitted] • Targeted metabolomics analysis for 15 carotenoid-related metabolites. • Acute cooling stress on oxycarotenoids of koi was reversible. • Thermal stress changed the skin lightness via the epinephrine-guanine pathway. • Oxycarotenoids might be favorable for enhancing koi skin color saturation. [ABSTRACT FROM AUTHOR]

Quality evaluation of koi (Cyprinus rubrofuscus) is essential to the koi industry. However, the community has a lack of knowledge about the quality evaluation of koi and about features of koi which are valuable. During the “All Japan Nishikigoi Show” competition, a quality evaluation of koi was conducted by authoritative experts. In the present study, photos of participants of this competition were used for analysis to discover features affecting the quality of the Kohaku variety of koi. The HSVA color model was used to define and extract from image red and pale red coverage rates in Kohaku body coloration. Body aspect ratio, body proportions, red coverage rate, and pale red coverage rate were extracted using tools written in Python programming language. Analyses of the extracted data revealed the significant impact body aspect ratio (p < 0.001) and pale red coverage rate (p < 0.001) on Kohaku quality. A low body aspect ratio has a positive impact on Kohaku quality, while a high pale red coverage rate has a very strong negative impact on Kohaku quality. This study and its results can be an important foundation for further research about koi quality.

Koi herpesvirus (KHV) has been identified in common carp (Cyprinus carpio) and varieties of C. carpio such as koi carp. Handling KHV outbreak on koi fish are always late and on case KHV infection, hence the purpose of this study is to find an alternative method in identifying a particular fish diseases KHV which more simple, practical and accurate. 20 samples are positively infected by KHV were derived from Blitar, with the characteristic of them are gill slime, erosion or ulcers on the skin, movement were not controlled, and skin blister. The result of our study showed that 100% positive reaction to the ICC in smear blood, which is marked golden chocolate color. The result of immunostaining streptavidin biotin obtained that koi fish have been infected by severe KHV showed golden chocolate color on the blood smear compared to positive control. While fish have been infected by undemanding KHV infection or still in the beginning stage, the blood smear showed chocolate bluish. It can be concluded that Immunocytochemistry Test Streptavidin Biotin can be used for early identifying of Koi Herpes Virus (KHV) due to simpler producer, more practical, and accurate result.

Objective: Present research aims to isolate, identify, and determine the virulence of the Streptococcus agalactiae (group B Streptococcus; GBS), isolated from popped eye disease affected Tilapia and Vietnamese Koi (V. Koi) fishes. Materials and Methods: A total of 330 fish samples were collected, of which Tilapia (n = 180) and V. Koi (n = 150), were collected from 35 affected ponds of four selected districts of Bangladesh. Isolation of the bacterium was done using different culture media (Nutrient broth, Plate count agar, Tryptic Soy Agar, and Blood agar), and identification by using various biochemical tests (conventional and using API 20 Strep kit) and polymerase chain reaction (PCR) using primers against 16S rRNA gene of S. agalactiae. Antibiotic susceptibility of the bacteria was performed using seven different antibiotics disc (Tetracycline, Oxytetracycline, Chlortetracycline, Streptomycin, Ciprofloxacin, Gentamicin, and Neomycin). Virulence of the isolated S. agalactiae was determined by infecting healthy Tilapia and V. Koi fishes through experimental infection. Results: Isolated bacteria were found Gram-positive paired and chained cocci, β-hemolytic and non-motile. Findings of biochemical and serological tests indicate that the isolated bacterium belongs to Group B Streptococcus of Lancefield classification. PCR result also confirmed that the bacteria were S. agalactiae. The bacterial isolates possessed resistance property against all the seven antibiotics used in this study. The isolated GBS was found highly virulent and showed 80%-90% mortality for Tilapia and V. Koi fishes in experimental infection within 1-6 days of post-infection. Conclusion: From the findings of this study, it may be concluded that isolated GBS from the Tilapia and V. Koi fishes were highly virulent and possessed multidrug-resistance properties. [ABSTRACT FROM AUTHOR]

Previously, a homodiploid goldfish-like fish (2 n = 100; GF-L) was spontaneously generated by self-crossing a homodiploid red crucian carp-like fish (2 n = 100; RCC-L), which was in turn produced via the distant hybridization of female koi carp (Cyprinus carpio haematopterus , KOC, 2 n = 100) and male blunt snout bream (Megalobrama amblycephala , BSB, 2 n = 48). The phenotypes and genotypes of RCC-L and GF-L differed from those of the parental species but were similar to diploid red crucian carp (2 n = 100; RCC) and goldfish (2 n = 100; GF), respectively. We sequenced the complete mitochondrial DNAs (mtDNAs) of the KOC, BSB, RCC-L, GF-L, and subsequent generations produced by self-crossing [the self-mating offspring of RCC-L (RCC-L-F2) to the self-mating offspring of RCC-L-F2 (RCC-L-F3) and the self-mating offspring of GF-L (GF-L-F2)]. Paternal mtDNA fragments were stably embedded in the mtDNAs of both lineages, forming chimeric DNA fragments. In addition to these chimeras, several nucleotide positions in the RCC-L and GF-L lineages differed from the parental bases, and were instead identical with RCC and GF, respectively. Moreover, RCC-L and GF-L mtDNA organization and nucleotide composition were more similar to those of RCC and GF, respectively, compared to parental mtDNA. Finally, phylogenetic analyses indicated that RCC-L and GF-L clustered with RCC and GF, not with the parental species. The molecular dating time shows that the divergence time of KOC and GF was about 21.26 Mya [95% highest posterior density (HPD): 24.41–16.67 Mya], which fell within the period of recent. The heritable chimeric DNA fragments and mutant loci identified in the mtDNA of the RCC-L and GF-L lineages provided important evidence that hybridizations might lead to changes in the mtDNA and the subsequent generation of new lineages. Our findings also demonstrated for the first time that the paternal mtDNA was transmitted into the mtDNA of homodiploid lineages (RCC-L and GF-L), which provided evidence that paternal DNA plays a role in inherited mtDNA. These evolutionary analyses in mtDNA suggest that GF might have diverged from RCC after RCC diverged from koi carp. [ABSTRACT FROM AUTHOR]

This is a proposal to encode the Old Permic script in the international character encoding standard Unicode. Old Permic was published in Unicode Standard version 7.0 in June 2014. The script was devised in the 14c to write the Komi and Komi-Permyak languages. It was used for Komi until the 17c. It was also used cryptographically from the 15c to write Russian.

Koi herpesvirus disease (KHVD) is a highly infectious disease leading to outbreaks and mass mortality in captive and free-ranging common carp and koi carp. Outbreaks may result in high morbidity and mortality which can have a severe economic impact along the supply chain. Currently, control and prevention of KHVD relies on avoiding exposure to the virus based on efficient hygiene and biosecurity measures. An early diagnosis of the disease is crucial to prevent its spread and to minimize economic losses. Therefore, an easy-to-handle, sensitive, specific and reliable test prototype for a point-of-care detection of KHV was developed and evaluated in this study. We used a multiplex-endpoint-PCR followed by a specific probe hybridization step. PCR-products/hybridization-products were visualized with a simple and universal lateral flow immunoassay (PCR-LFA). Fifty-four gill tissue samples (KHV-positive n = 33, KHV-negative n = 21) and 46 kidney samples (KHV-positive n = 24, KHV-negative n = 22) were used to determine diagnostic sensitivity and specificity of the PCR-LFA. In addition, the usability of PCR-LFA to detect CyHV-3-DNA in gill swabs taken from 20 perished common carp during a KHVD-outbreak in a commercial carp stock was examined. This assay gave test results within approximately 60 min. It revealed a detection limit of 9 KHV gene copies/μl (95% probability), a diagnostic specificity of 100%, and diagnostic sensitivity of 94.81% if samples were tested in a single test run only. PCR inhibition was noticed when examining gill swab samples without preceding extraction of DNA or sample dilution. Test sensitivity coud be enhanced by examining samples in five replicates. Overall, our PCR-LFA proved to be a specific, easy-to-use and time-saving point-of-care-compatible test for the detection of KHV-DNA. Regarding gill swab samples, further test series using a higher number of clinical samples should be analyzed to confirm the number of replicates and the sample processing necessary to reveal a 100% diagnostic sensitivity. [ABSTRACT FROM AUTHOR]

CASE DESCRIPTION A client-owned 12-year-old 10.3-kg (22.7-lb) female shusui koi (Cyprinus carpio) was evaluated because of an ulcerated mass on the left body wall, hyporexia, and decreased activity. CLINICAL FINDINGS The patient was anesthetized with a solution of eugenol in water for all examinations and procedures. An approximately 7 × 5-cm smooth, raised, ulcerated, and firm mass was present ventral and lateral to the dorsal fin on the left body wall. Whole-body CT images obtained before and after contrast administration revealed an encapsulated, homogeneous, fat-opaque mass within the muscle. The mass was fat echoic with poor vascularity on ultrasonographic examination. Histologic evaluation of an ultrasound-guided needle biopsy specimen was suggestive of a lipoma. TREATMENT AND OUTCOME The mass was excised, and the fish was placed in water with 0.3% salinity for 3 weeks after surgery. Postoperative antimicrobial administration was not indicated, and additional postoperative analgesic administration was considered impractical. The patient had noticeable improvement in appetite and activity with no indication of discomfort immediately following surgery. Five weeks after surgery, the incision site had healed with minimal scarring, and evaluation of CT images revealed no evidence of mass regrowth or regional osteomyelitis. CLINICAL RELEVANCE Antemortem evaluation and diagnosis of a lipoma in a teleost with subsequent excision was described. This report highlighted the logistic challenges associated with anesthesia, advanced diagnostic imaging, and surgery in fish and showed that they can be successfully overcome so that high-level medical care can be provided to such patients. [ABSTRACT FROM AUTHOR]

The present study systematically investigated the effects of warehouse and greenhouse aquaponic growing conditions on consumer acceptability of different basil cultivars. A total of 105 consumers rated their liking of three basil cultivars (Nufar, Genovese, and Eleonora), each grown in three conditions (aquaponically in a greenhouse, aquaponically in a warehouse, both with Cyprinus carpio, Koi fish, and grown in soilless medium). We used linear random effect models to investigate consumer preferences for attributes of basil plants grown in different environments by controlling for individualspecific random effects. Participants generally liked the soilless medium-grown and greenhouse aquaponically grown basil plants more than the warehouse aquaponically grown plants. The soilless medium-grown basil had the highest appearance liking and flavor intensity, followed by the greenhouse aquaponic grown and then by the warehouse aquaponic grown. Aquaponically grown cultivars were rated less bitter than soilless medium-grown cultivars. [ABSTRACT FROM AUTHOR]