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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]

Carp edema virus (CEV) is the causative agent of carp edema virus disease (CEVD), also known as koi sleepy disease (KSD), which is a highly pathogenic viral infection of koi and common carp. Initially identified in Japan in 1974, the spread of CEVD has escalated in recent years with the increasing global trade of koi, and has been documented throughout Europe and Asia, thereby negatively impacting koi aquaculture. Between 2021 and 2022, an infectious disease outbreak occurred in koi farms across various regions of Beijing. Following clinical diagnosis, histopathological examination, and molecular testing, CEV was identified as the causative agent of the epidemics. Using the detection method designed by the Centre for Environment, Fisheries and Aquaculture Science(CEFAS), 11 positive samples were detected by PCR, including nine that were verified by nested PCR. Subsequently, nested PCR and phylogenetic analysis confirmed that CEV was the main pathogen of koi farms. Additionally, increased water temperature was positively correlated to the mortality rate of CEV-infected koi, indicating that temperature is an important factor influencing the incidence of CEV infection. The findings of this study provide valuable references for the development of new strategies for the diagnosis and prevention of CEVD. [ABSTRACT FROM AUTHOR]

The objective of this study was to examine the effectiveness of using ultrasonography for sex determination in young carp. Ultrasound images were obtained from anesthetized captive carp aged 11 to 13 months. After sacrifice, the body length and weight, gonadal sex and weight, and gonad-somatic index were measured. Well-developed testes and ovaries filled with granulated eggs were easily distinguishable on ultrasound imaging, based on their visible characteristic morphological features. In contrast, immature, small-sized testes and ovaries lacking granulated eggs were difficult to distinguish. The mean longitudinal thickness of gonads was technically the easiest to scan for measuring gonadal size, and it correlated well with gonadal weight (r = 0.91) and gonad-somatic index (r = 0.89). Based on the area under the curve of the receiver operating characteristic curve, we determined that 6.2 mm was the cutoff value for mean longitudinal gonadal thickness at which sex could be positively determined by ultrasonography, with a sensitivity of 82.6% and specificity of 100%. Ultrasonography therefore represents an effective and non-invasive diagnostic tool for determining sex and maturity in Japanese ornamental carp. [ABSTRACT FROM AUTHOR]

The presence of conspecifics plays a crucial role in mitigating stress responses in social teleost species and holds potential for enhancing welfare in captive fish. While studies on social buffering effects have typically focussed on single species, marked interspecific differences can exist. Here, we conducted an analysis of social buffering of stress response in two of the most extensively farmed fish, the Nile tilapia and the koi carp. Subjects were exposed to a behavioural stress response assay (open‐field test) in three conditions simulating increasing levels of social enrichment: isolation, pairs, or shoals of five fish. We obtained five stress indicators from the assay: thigmotaxis, freezing, activity, erratic movements and interindividual distance in conditions with more than one fish. In both species, erratic movements significantly decreased with increasing levels of social enrichment, suggesting a similar social buffering effect. However, other indicators revealed species differences. Koi carp, but not Nile tilapia, showed a socially‐mediated reduction in thigmotaxis, whereas Nile tilapia, but not Koi carp, showed a socially‐mediated reduction in freezing behaviour. Furthermore, social enrichment determined opposite effects on the activity of the two species: Nile tilapia were more active as group size increased, whereas the opposite trend was found in koi carp. Finally, Nile tilapia showed increased interindividual distance with increasing social group size, whereas no changes were observed for koi carp. Our study indicates that the buffering effects of social enrichment on the behavioural stress response do not completely overlap between different fish species, highlighting the importance of developing finely‐tuned species‐specific enrichments and welfare indicators. [ABSTRACT FROM AUTHOR]

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. [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]

Captive fish populations, such as those encompassing aquarium and pet fish, offer significant economic value and are integral to conservation, research, and education. However, these ornamental fish exhibit a reduced ability to protect their ocular surfaces, and our understanding of the ocular diseases that affect them remains limited. Although corneal neoplasms in carp are uncommon, identifying their distinct characteristics is crucial in selecting appropriate therapeutic interventions that aim to preserve vision, prevent the ocular loss, and ultimately ensure the survival of the affected fish. This study provides clinical and histopathological details of various proliferative corneal masses in Cyprininae species, including five koi (Cyprinus carpio) and four goldfish (Carassius auratus). It discusses a spectrum of neoplasms, including soft tissue sarcoma, spindle cell sarcoma, chromatophoroma, and papilloma, in addition to conditions like exuberant granulation tissue and proliferative carp pox. These findings bear significant implications for clinical decision‐making and treatment, offering valuable insights into the incidence and characteristics of corneal tumors in captive fish, which could inform further studies in this area. [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]

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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]

Koi is a brightly coloured, elegant fish with high ornamental value, and is known as a "swimming work of art". Fish body colour is mainly determined by the type and distribution of pigment cells on the skin or scales. In this paper, the composition, distribution and morphology of pigment cells in the scales and skin of four different body colours, namely, black, white, yellow and red koi, were compared. The composition of pigment cells in the scales and skin was consistent, and rich iridescent cells were present in all four koi, with melanophores in black koi, and xanthophores and erythrophores in yellow and red koi. Only iridophores were observed in white koi. The ultrastructure of iridophores in all four koi colours was observed by transmission microscopy, and the iridophores mainly had three different morphologies: pike-shaped, short rod-shaped and vacuolated. Real-time fluorescence quantitative PCR was used to detect the expression of eight iridocyte development-related genes in the skin of four different body colors of koi carp, the results showed that eight genes related to iridophores development were expressed in the skin of different body colours, whereas there were differences in expression, among which the relative expression of alk, pka, sox10, pax3, foxd3 and pnp4a genes was higher in red and yellow koi, while the relative expression of ltk and alx4a genes was the highest in white koi. In order to further confirm the role of the pnp4a genes in iridophores development, the recombinant plasmid of overexpression of pnp4a gene was constructed, and was microinjected into the fertilized eggs of AB zebrafish. In order to further confirm the role of pnp4a gene in fish body colour formation, especially in iridophores, we constructed a pnp4a overexpression recombinant plasmid and microinjected it into the fertilized eggs of zebrafish (AB). [ABSTRACT FROM AUTHOR]

Fish's skin serves a variety of functions that are essential for survival, including communication, respiration, sensory perception, excretion, ion regulation, and heat regulation. This research aimed to examine the snout skin of koi fish (Cyprinus carpio) to determine its structural characteristics. Using light microscopy, the histochemical elements of the skin were examined in a total of 20 adult, healthy koi fish. The skin of was made up of three layers: epidermis, dermis, and hypodermis. The epidermis consisted of epidermal cells, club cells, mucous goblet cells, rodlet cells, eosinophilic granular cells, serous goblet cells, and melanocytes. large amounts mucous of cells which reacted positive to Alcian blue (AB) and Periodic Acid-Schiff (PAS), few club cells which reacted positive to bromophenol blue and light green. many eosinophilic granular cells (EGCs) reacted positive to PAS and light green. Moreover, many taste buds also were demonstrated. The dermis was consisted of collagenous bundles, bundles of myelinated nerve fibers, blood vessels and immune cells. Many telocytes were detected in dermis in close relation to nerve fibers, blood vessels and immune cells. The lateral line system was well-developed in koi fish, where it was arranged into mechanoreceptive superficial and canal neuromasts and electroreceptive tuberous and ampullary organs. In conclusion, the epidermis of snout region in Koi fish consisted of a variety of cells with diverse functions. [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 document is a collection of poems titled "Koi Variations," "What I wrote in the water," "Lake Calm," "Eclipse," and "Entropy." The poems explore themes of nature, love, and the passage of time. "Koi Variations" depicts the movements of a koi fish and the moon, while "What I wrote in the water" reflects on love and memories. "Lake Calm" describes still water and its origins, "Eclipse" recounts a personal experience of witnessing a lunar eclipse, and "Entropy" contemplates the transient nature of life and love. The poems offer a unique perspective on these subjects and invite readers to reflect on their own experiences. [Extracted from the article]

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.

Lack of mature male broodfish, insufficient sperm, and non-synchronized maturation times have always been a hindrance to the breeding program of Japanese koi (Cyprinus carpio var. koi) raised indoors. Therefore, it is believed that the preservation of Japanese koi sperm by short-term storage and cryopreservation could solve this problem. In this study, the appropriate diluent solution, sperm-to-diluent ratio, and storage temperature for short-term storage of Japanese koi sperm were determined, and the efficacy of the short-term stored sperm in fertilizing eggs was evaluated. Milt samples collected from sexually mature males were pooled and tested in modified calcium-free Hank's Balanced Salt Solution (CF-HBSS), modified Mahseer extender, and modified Kurokura extender at 1:1 and 1:5 ratios of sperm to diluent, respectively. Storage temperatures were tested at 4 °C and room temperature. Milt sample without diluent solution served as a control. The percentage of sperm motility was measured daily for one week. For the egg fertilization experiment, Japanese koi eggs were fertilized with sperm on the second day of short-term storage, while a freshly collected sperm sample served as a control. We found that sperm diluted 1:1 with a modified Kurokura extender and stored at 4 °C had a mean sperm motility of 76.00 ± 3.06% on the third day, compared with 54.67 ± 2.91% in the control treatment (P<0.05). Short-term stored spermatozoa showed equivalent egg fertilization ability compared to fresh spermatozoa (control) (P>0.05). In conclusion, the use of a modified Kurokura extender at a 1:1 ratio of sperm to diluent and storage at 4 °C was optimal for short-term storage of Japanese koi sperm, and these sperm still showed equivalent egg fertilization ability to freshly collected sperm after two days of storage. In addition, the current study also determined the appropriate extender solution for cryopreservation of Japanese koi sperm. [ABSTRACT FROM AUTHOR]

Chinese giant salamander iridovirus (GSIV) is the first known and causative viral pathogen in Andrias davidianus. Developing a sensitive, accurate and specific assay to detect GSIV in samples is essential to prevent the further spread of the pathogen. In this study, we established a droplet digital PCR (ddPCR) assay that targeted the mcp gene of GSIV, enabling rapid and quantitative detection of the virus. We determined that the optimal annealing temperature, primer concentration and probe concentration were 57.1°C, 50 nM and 500 nM, respectively. We analysed the specificity and sensitivity of the ddPCR assay and found that five common aquatic animal viruses, including Cyprinid herpesvirus 2 (CyHV‐2), infectious spleen and kidney necrosis virus (ISKNV), Koi herpesvirus (KHV) and Carp Edema Virus (CEV) displayed negative results based on this GSIV ddPCR assay. The assay can detect GSIV with the lowest detection limit of 3.7 copies per reaction. To evaluate the sensitivity and accuracy of the ddPCR assay, we tested different infected tissue samples with both the ddPCR and TaqMan real‐time PCR assays. Our results showed that the ddPCR assay detected GSIV in all samples with 100% positivity, while the TaqMan real‐time PCR assay detected GSIV in only 82.1% of samples. The established ddPCR method provided several advantages in detecting GISV, including high sensitivity, high precision and absolute quantification, making it a powerful tool for detection of possible and potential GSIV infection, even in samples with low viral load. [ABSTRACT FROM AUTHOR]

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]

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. • The immune responses of koi carp to A. hydrophila were described for the first time. • 111,650 unigenes and 252 DEGs were obtained. • Key genes and pathways response to A. hydrophila were identified. [ABSTRACT FROM AUTHOR]

Cyprinid herpesvirus 3 (CyHV‐3) is the main pathogen of koi herpesvirus disease (KHVD), which has caused serious damage to the ornamental and food‐producing carp industry. Effective and rapid on‐site detection methods are needed for early diagnosis of CyHV‐3. A lateral flow immuno‐chromatographic assay (LFIA) using two specific anti‐CyHV‐3 monoclonal antibodies has been developed and validated for on‐site detection of CyHV‐3. MAb 3C9 was used to bio‐conjugate CyHV‐3 antigen with colloidal gold, and MAb 2A8 was used to capture antigen bound colloidal gold on the test line. The control line was lined with goat anti‐mouse IgG to capture unbound colloidal gold to validate performance. The test results can be viewed within 10 min after putting the strip into CyHV‐3 virus infection fluid. The lowest limit of detection for the LFIA test was found to be 1.5 × 104 copies/μL and it showed no cross‐reactivity with other fish viral pathogens. The specificity of the strip was 100% when spleen and kidney tissues of CyHV‐3‐infected and healthy koi were validated at the field level. The LFIA strip will be an effective device for the early detection of CyHV‐3 in the future. [ABSTRACT FROM AUTHOR]

Background: One cause of fish disease is the Argulus parasite. Both male and female Argulus were found to infect fish. This study aims to determine the percentage difference of male and female Argulus that infect Cyprinidae in Magelang Regency. Methods: The present study used a survey method. The independent variables in this study are carp (Cyprinus carpio), goldfish (Carassius auratus), comet goldfish (Carassius auratus), koi fish (C. carpio koi) and the sex of the Argulus parasite. The dependent variable of this study was the sex percentage of the Argulus parasite. The data analysis was done using analysis of variance (ANOVA) then followed by Duncan's multiple range test to find out the differences between treatments. Result: The first ANOVA result showed no significant difference (p>0.05), the second ANOVA showed significantly different results (p<0.05) and the third ANOVA reveals no significantly different percentage (p>0.05). The highest infestation rate of male Argulus japonicus is found in koi fish (C. carpio koi) is 60% and the lowest is in comet goldfish, which is 38.46%. Whereas infestation of female A. japonicus in carp, goldfish, comet goldfish and koi fish also obtained a similar result. [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) is the causative agent of a koi herpesvirus disease (KHVD) inducing high mortality rates in common carp and koi (Cyprinus carpio). No widespread effective vaccination strategy has been implemented yet, which is partly due to side effects of the immunized fish. In this study, we present an evaluation of the purification of infectious KHV from host cell protein and DNA, using the steric exclusion chromatography. The method is related to conventional polyethylene glycol (PEG) precipitation implemented in a chromatographic set‐up and has been applied for infectious virus particle purification with high recoveries and impurity removal. Here, we achieved a yield of up to 55% of infectious KHV by using 12% PEG (molecular weight of 6 kDa) at pH 7.0. The recoveries were higher when using chromatographic cellulose membranes with 3–5 μm pores in diameter instead of 1 μm. The losses were assumed to originate from dense KHV precipitates retained on the membranes. Additionally, the use of >0.6 M NaCl was shown to inactivate infectious KHV. In summary, we propose a first step towards a purification procedure for infectious KHV with a possible implementation in fish vaccine manufacturing. [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]

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. [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]

Specific viruses were additionally examined via conventional PCR, with partial SphI-5 (610 bp), 9/5 (611 bp) (Kurita et al., [7]) and thymidine kinase genes (1117 bp) amplified (Padhi et al., [10]; Appendix S3). While the 2018-2019 viruses from the Pardubice region were 100% identical, the CyHV3-J virus CZ2296 was significantly variable (9 bp difference) and the virus CZ2300 collected in 2020 showed 1 bp variability, along with virus CZ2241 originating from outside of the Pardubice region. Latent infection, PCR, Sanger sequencing, thymidine kinase, virus transmission Keywords: latent infection; PCR; Sanger sequencing; thymidine kinase; virus transmission EN latent infection PCR Sanger sequencing thymidine kinase virus transmission 709 713 5 05/09/23 20230601 NES 230601 INTRODUCTION Cyprinid herpesvirus 3 (koi herpesvirus, KHV) caused a number of outbreaks in the Pardubice region (Czech Republic) in the 2018-2020 period. [Extracted from the article]

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]

In order to investigate the genetic diversity and variation of different body colors of common carp (Cyprinus carpio), a total of 150 individuals from six different colored koi carp populations (Platinum Ogon, Higoi, Kohaku, Tajsho-sanke, Shiro Utsuri and Hi Utsuri), two colored FFRC No.2 strain common carp (grey and red) populations, and two hybrid carp populations (grey FFRC No.2 strain common carp♀ × Kohaku koi♂,Kohaku koi♀ × grey FFRC No.2 strain common carp♂) were analyzed based on mtDNA D-loop sequences. The results showed that the contents of A, T, G and C in the 932 bp D-loop sequence were 33.2%, 32.8%, 14.2% and 19.8%, respectively. In total, 34 variant sites were detected in all individuals, showing 18 haplotypes. Among them, there were two dominant haplotypes in Kohaku and Hi Utsuri, respectively. There were two unique haplotypes in white koi carp and grey FFRC No.2 strain common carp, and grey and red FFRC No.2 strain common carp shared one dominant haplotype. The genetic distances across the ten populations ranged from 0.001 to 0.018, with FST values ranging from 0.011 to 0.933. The analysis of molecular variance (AMOVA) revealed that there was 67.17% variation among varieties and 32.83% variation within varieties, indicating highly substantial genetic difference. It demonstrated that the Hi Utsuri, white koi carp, and Tajsho-sanke had high genetic diversity, while the Shiro Utsuri, FFRC No.2 strain common carp (grey and red), and hybrid carp had high genetic purity. The study enriched the genetic data of the various body color carp germplasm resources and provided the guidance for germplasm resource utilization and genetic selection in the future. [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]

Objective: A privately owned, 4‐year‐old female Showa koi (ornamental variant of Common Carp Cyprinus carpio) was presented for erratic swimming, air gasping, water spitting, and abnormal orientation in the water column. Initial physical examination revealed an obese patient with no external abnormalities except a small plaque localized to filaments on a right gill hemibranch. Methods: The fish was anesthetized using AQUI‐S 20E (10% eugenol solution) at 50 mg/L to facilitate diagnostic evaluation. Biopsy of the gill lesion yielded no significant findings. Whole‐body computed tomography confirmed an excess of adipose tissue and mild scoliosis, with narrowing of the 10th–11th intervertebral space. A weight loss plan and need for repeat assessment were recommended. Result: The patient's original abnormal behaviors resolved over the following weeks, but it subsequently became hyporexic and depressed. Full‐body magnetic resonance imaging (MRI) showed extensive alteration of vertebral centra, with multifocal compression of the spinal cord. Due to the patient's declining clinical condition and the grave prognosis based on MRI findings, the patient was humanely euthanized. Conclusion: Postmortem examination showed severe transmural myelomalacia associated with a vertebral subluxation. This case demonstrates the practical application of advanced cross‐sectional imaging that has not been commonly afforded to fish or other lower vertebrates. Impact StatementThough magnetic resonance imaging (MRI) is a key part of evaluating patients for diseases of the nervous system, it has largely only been utilized in fish in research contexts. This paper puts forth one of the first usages of magnetic resonance imaging in a fish in a clinical setting. [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]

The purpose of the study was to investigate the inheritance and expression of the koi color dominant mutations "design" and "red eyes" in koi × Goldfish hybrids (koi Cyprinus carpio × Goldfish Carassius auratus). The mutation "design" manifests as a yellow stripe along the dorsal fin and a specific ornament on the head; this trait is controlled by a dominant allele of one gene (D/d). In four progenies that were obtained by crosses of koi females having the "design" trait with Goldfish males, the segregations of so‐called "ghost fish," having a melanin‐pigmented body with the "design" pattern, and fish having wild‐type color, without the "design" pattern, were close to the Mendelian 1:1 ratio. This indicates that koi females with the "design" trait that are used for the production of these progenies had the genotype Dd, whereas the Goldfish males had the genotype dd. In one progeny that was obtained by crossing a koi female having the "design" trait with a Goldfish male, 420 ghost fish (99.8%) and one fish with wild‐type color (0.2%) were recorded. The koi female from which this progeny originated was apparently homozygous for the dominant allele (genotype DD). Earlier, it was shown that the appearance of red eyes in koi is caused not by the albino mutation, but by another dominant demelanization mutation (R). In three progenies that were obtained by crossing koi females having red eyes with Goldfish males, the segregations of the larvae with unpigmented bodies and light eye lenses and those with dark bodies and black eye lenses were close to 1:1. Later, three types of body color were observed in the juveniles of these progenies: yellow, brown, and wild‐type; all of the juveniles had black eyes. The segregations of colored (yellow + brown) and wild‐type‐color juveniles were close to 1:1. This shows that that the red‐eyed koi females that were used in the crosses were heterozygous (genotype Rr), whereas the Goldfish males had genotype rr. The described colored forms of koi × Goldfish hybrids can be used for weed control. [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]

The aim of the present study was to optimize the solid state co‐fermentation requirements of soybean meal (SBM) by Bacillus subtilis and B. licheniformis and assess the application effects on the feeding of koi carp. The soluble protein conversion rate (SPCR) was the highest when co‐fermented by B. subtilis and B. licheniformis with the mixed ratio 1:2 (V/V, marked as SL1:2). As the results of response of surface methodology showed, the optimal co‐fermentation requirements by SL1:2 were at inoculums size 5% under the duration of 60 h at 30°C and with the substrate/water ratio 1:1. The highest SPCR was 50.22% ± 2.06%, and the anti‐nutrient factors in fermented soybean meal (FSBM) decreased significantly (p < 0.05). Six isonitrogenous diets were formulated. Fish meal (FM) was partially replaced by 0%, 30% and 60% of FSBM in the treatment groups, and FSBM replaced by SBM as controlled group. When FM was replaced by 0% and 30% of FSBM, the effects of FSBM on protease activity, immune activity and antioxidant activity of Koi were higher than that of the same proportion of SBM. Meanwhile, the effects of FSBM on the feeding of koi carp could reduce the dosage of 10% SBM at the same proportion. As the results show, FSBM used in aquaculture effectively reduced feed expenditure, improved economic and environmental benefits, and prevented the deterioration of water quality. [ABSTRACT FROM AUTHOR]