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Heavy metal contamination of aquatic environments adversely affects the health of aquatic organisms and consumption of fish contaminated with heavy metals poses serious health risks to humans. Among various strategies, probiotics (living microorganisms known to have beneficial effects on the host), which have been extensively applied in the aquaculture industry, could be helpful for heavy metal detoxification and remediation. Several probiotics, including Lactobacillus strains, exhibit heavy metal binding, high heavy metal tolerance, and other beneficial characteristics for the host. Notably, numerous probiotics have been reported to bind heavy metals and excrete them from the host. Various probiotic strains (Lactobacillus, Bacillus, Lactococcus, etc.) show beneficial effects in alleviating heavy metal toxicity in cultured fish species. Certain probiotic bacteria reduce the absorption and bioavailability of heavy metals by enhancing heavy metal detoxification and sequestration while preserving gut barrier function. This review summarises the toxic effects of selected heavy metals on the health of farmed fish and discusses the role of probiotic strains in remediating the consequential exposure-induced immune toxicity and oxidative stress. Moreover, we discussed the protective strategies of probiotics against heavy metal accumulation in various tissues and gut dysbiosis in fish to alleviate heavy metal toxicity in fish farming, thereby promoting a sustainable blue economy worldwide., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Se Chang Park reports article publishing charge was provided by Companion Animal Research of the Rural Development Administration, Republic of Korea., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

A female narrow-ridged finless porpoise ( Neophocaena asiaeorientalis ) stranded on a beach on Jeju Island showed epithelial proliferative skin lesions on its body. Two false killer whales ( Pseudorca crassidens ), caught using nets near Gangneung and Samcheok, respectively, had multiple plaques on their penile epidermis. Histological examination of the epidermis revealed that all the lesions had common features, including accentuated rete pegs, ballooning changes, and eosinophilic intranuclear inclusion (INI) bodies. Based on the histopathological results, herpesvirus infection was suspected, and thus further analysis was conducted using herpesvirus-specific primers. Based on nested polymerase chain reaction (PCR) tests using the herpesvirus-detectable primers, the PCR products demonstrated two fragments: a 222-base-pair (bp) sequence of the DNA polymerase gene, SNUABM_CeHV01, showing 96.4% identity with a bottlenose dolphin herpesvirus from the Jeju narrow-ridged finless porpoise; and a 222 bp sequence of the DNA polymerase gene, SNUABM_CeHV02, showing 95.95% identity with the same bottlenose dolphin herpesvirus from the Gangneung and Samcheok false killer whales. The significance of this study lies in its ability to demonstrate the existence of novel cetacean herpesviruses in South Korean seawater, representing an important step forward in studying potentially harmful pathogens that affect endangered whale and dolphin populations.

Despite the precarious state of marine turtles as a highly endangered species, our understanding of their diseases remains limited. This case report presents a detailed pathological investigation of spirorchiidiasis, a blood fluke infection that poses a substantial threat to marine turtles. This retrospective study examined three cases of spirorchiid-infected sea turtles, specifically, green sea turtles stranded on Jeju Island, South Korea. Premortem examination of the three spirorchiid-infected green sea turtles demonstrated nonspecific clinical symptoms; blood analysis revealed dehydration, malnutrition, and anemia. Computed tomography scans provided insights into severe pulmonary and extrapulmonary manifestations, including the mass present in the joint region. Post-mortem examinations consistently indicated severe lung lesions and systemic manifestations, with histopathological examination confirming the presence of spirorchiid ova across various organs. Despite the global prevalence of spirorchiidiasis in sea turtles, disease severity varies regionally. This report provides a detailed demonstration of the pathology of spirorchiidiasis in sea turtles from Northeast Asia.

Erwinia amylovora, the primary causative agent of blight disease in rosaceous plants, poses a significant threat to agricultural yield worldwide, with limited effective countermeasures. The emergence of sustainable alternative agents such as bacteriophages is a promising solution for fire blight that specifically targets Erwinia. In this study, we isolated pEp_SNUABM_01 and pEa_SNUABM_55 from a South Korean apple orchard soil, analyzed their genomic DNA sequences, and performed a comprehensive comparative analysis of Hena1 in four distinct sections. This study aimed to unveil distinctive features of these phages, with a focus on host recognition, which will provide valuable insights into the evolution and characteristics of Henunavirus bacteriophages that infect plant pathogenic Erwinia spp. By elucidating the distinct genomic features of these phages, particularly in terms of host recognition, this study lays a foundation for their potential application in mitigating the risks associated with fire blight in Rosaceae plants on a global scale., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Heat-killed probiotics offer an alternative approach to enhance growth and disease resistance in farmed fish. In this study, we isolated Lactiplantibacillus plantarum VSG3 from the gut of Labeo rohita to investigate the effects of heat-killed L. plantarum (HK-LP) on the health and growth performance of Cyprinus carpio fingerlings. Different concentrations of HK-LP (0, 50, 100, 200, 300, and 400 mg/kg) were administered to the fish, followed by a challenge with Aeromonas hydrophila after 8 weeks of feeding. Notably, the LP200 group exhibited significantly improved percentage weight gain and specific growth rate, accompanied by the lowest feed conversion ratio. Post-challenge survival rates were considerably enhanced in the LP200 group, reaching 60.65%. Moreover, serum analysis indicated significantly higher levels of total protein and albumin in the LP200 group than in the control group. Although HK-LP had no substantial impact on certain serum parameters (glucose, total cholesterol, cortisol, and alanine aminotransferase), aspartate aminotransferase levels were considerably low in the LP200 group. Intestinal protease and trypsin activities significantly increased in the LP200 group, while no significant changes were observed in lipase and amylase activities post-pathogen challenge. Serum immunological indices, including lysozyme, alternative complement pathway, and phagocytic activity, improved considerably in the LP200 group. Additionally, serum antioxidant enzyme activities (superoxide dismutase [SOD], glutathione peroxidase [GPx], catalase [CAT], and myeloperoxidase) were significantly elevated in the LP200 group, while malondialdehyde level was reduced. Gene expression analysis in liver tissue indicated strong upregulation of antioxidant-related genes (SOD, CAT, nuclear factor erythroid 2 [NFE2]-related factor 2 [Nrf2], Kelch-like ECH-associated protein 1[Keap1]) in the LP100 and LP200 groups. Pro-inflammatory cytokines (IL-1β and TNF-α) were considerably downregulated in the kidneys of the LP200 post-challenged fish, although the anti-inflammatory cytokine IL-10 showed an increased expression. Quadratic regression analysis identified the optimal dietary HK-LP level for maximizing growth and immune performance (200.381-270.003 mg/kg). In summary, our findings underscore the potential of HK-LP as a valuable dietary supplement for enhancing carp aquaculture, particularly at the appropriate concentration., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Human fishing activities have significantly affect environmental concern for marine ecosystems, conservation of marine mammals, and human health. Coastal cetaceans are highly vulnerable to ingestion of fishing gear, bycatching, or entanglement, all of which can be fatal for these animals. In particular, certain coastal dolphins and porpoises are heavily impacted by fishing gear such as angling gear or stownet, as their food often overlap with the target fish species of human fisheries., Case Presentation: This study presents a case of an Indo-Pacific finless porpoise (Neophocaena phocaenoides) beached on the coast of Jeju Island, Republic of Korea, with ingestion of fishing gear and severe Anisakis infection. Although this species inhabits waters ranging from the Persian Gulf to Taiwan, several stranded carcasses have been reported on Jeju Island in recent years. Post-mortem computed tomography revealed a bundle of four fishing hooks in the forestomach, along with nylon lines and steel lines with connectors, which were assumed to be angling gear for Jeju hairtail (Trichiurus lepturus). Further necroscopic investigation revealed that the forestomach contained a large number of Anisakis spp. (Nematoda: Anisakidae). Histological examination revealed a thickened forestomach wall with pinpoint and volcanic ulcerations, a thickened layer of stratified squamous epithelium, and infiltrated stroma in the squamous epithelium., Conclusions: This study emphasizes the urgent need to address the impact of fishing activities on marine mammals, marine litter pollution, and the bycatch problem in Korean seawater. In addition, the occurrence of N. phocaenoides in seawater around Jeju Island should be raised in future geographical ecology or veterinary pathology studies and when its distribution is updated., (© 2024. The Author(s).)

The pancreas is a heterocrine gland that has both exocrine and endocrine parts. Most pancreatic cancer begins in the cells that line the ducts of the pancreas and is called pancreatic ductal adenocarcinoma (PDAC). PDAC is the most encountered pancreatic cancer type. One of the most important characteristic features of PDAC is neuropathy which is primarily due to perineural invasion (PNI). PNI develops tumor microenvironment which includes overexpression of fibroblasts cells, macrophages, as well as angiogenesis which can be responsible for neuropathy pain. In tumor microenvironment inactive fibroblasts are converted into an active form that is cancer-associated fibroblasts (CAFs). Neurotrophins they also increase the level of Substance P, calcitonin gene-related peptide which is also involved in pain. Matrix metalloproteases are the zinc-associated proteases enzymes which activates proinflammatory interleukin-1β into its activated form and are responsible for release and activation of Substance P which is responsible for neuropathic pain by transmitting pain signal via dorsal root ganglion. All the molecules and their role in being responsible for neuropathic pain are described below., (© 2024 Wiley Periodicals LLC.)

Introduction: Spinal deformities, including kyphoscoliosis, have been consistently documented in cetaceans. However, the majority of reported cases of kyphoscoliosis in cetaceans pertain to bottlenose dolphins, with limited information on its occurrence in narrow-ridged finless porpoise (NFP) (Neophocaena asiaeorientalis)., Materials and Methods: In November 2021, two deceased NFPs were discovered stranded on the shores of the Republic of Korea. As part of the pioneer stranded cetacean imaging programme in the Republic of Korea, both carcasses underwent post-mortem computed tomography (PMCT), revealing congenital and degenerative traumatic kyphoscoliosis, respectively., Results: Although kyphoscoliosis may not have directly caused the demise of these individuals, it is hypothesized that the reduced spinal range of motion and mobility associated with kyphoscoliosis may have contributed to their deaths., Conclusion: This case report presents the first documented cases of kyphoscoliosis in two NFPs stranded in Korean waters, utilizing PMCT as an efficient methodology for assessing skeletal abnormalities in cetaceans., (© 2024 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Fire blight is a bacterial disease that affects plants of the Rosaceae family and causes significant economic losses worldwide. Although antibiotics have been used to control the disease, concerns about their environmental impact and the potential to promote antibiotic resistance have arisen. Bacteriophages are being investigated as an alternative to antibiotics; however, their efficacy can be affected by environmental stresses, such as UV radiation. In this study, we optimized the formulation of Erwinia phages to enhance their stability in the field, focusing on improving their UV stability and adsorption using adjuvants. Our results confirmed that 4.5 % polysorbate 80 and kaolin improve phage stability under UV stress, resulting in an 80 % increase in PFU value and improved UV protection efficacy. Adsorption assays also demonstrated that polysorbate 80 and kaolin improved the absorption efficiency, with phages detected in plant for up to two weeks. These findings demonstrate the effectiveness of the auxiliary formulation of Erwinia bacteriophages against environmental stress., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

This study investigated the effects of dietary piperine (PIP) on growth performance, digestive enzymes, serum biochemical parameters, antioxidant and immune responses, and gene expression in Cyprinus carpio challenged with Aeromonas hydrophila. Six diets were prepared with PIP doses of 0, 0.5, 1.0, 2.0, 3.0, and 4.0 g/kg, corresponding with the control, PR50, PR100, PR200, PR300, and PR400, respectively. Fish were challenged with Aeromonas hydrophila after 8 weeks of feeding with the respective diets. Weight gain (PWG) and specific growth rate (SGR) were significantly enhanced, whereas feed conversion ratio (FCR) was lowered in PR200. The cumulative post-challenge survival was improved to 68.43% in the PR200 group compared with 28.08% in the control. Serum total protein and albumin levels were significantly enhanced in the PR200 group compared to the control. However, dietary PIP up to 3 g/kg had no significant effect on serum glucose, cortisol, aspartate aminotransferase, or alkaline phosphatase activities; however, the alanine aminotransferase level was lower (P < 0.05) in the PR200 group than in the control. Intestinal amylase, lipase, and protease activities increased in PR300, and intestinal amylase and lipase increased in the PR100 group (P < 0.05). The serum immunological indices (lysozyme, alternative complement pathway, phagocytic activity, and respiratory burst activity) were higher (P < 0.05) in the PR200 group than in the control group. Serum superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities were significantly intensified in PR200-PR300 than in the control group, with the highest activity observed in the PR200 group. Malondialdehyde was significantly lower in the PR200 group than in the control group. Furthermore, SOD, CAT, and Nrf2 expression was strongly upregulated in the liver tissue of the PR200 and PR300 groups compared to that in the control. The transcript levels of pro-inflammatory cytokines viz. IL-1β and TNF-α were significantly upregulated in the kidneys of the PR100 and PR200 post-challenged. In contrast, the anti-inflammatory cytokine IL-10 was significantly downregulated in the kidneys of PR200. The expression of HSP70 was upregulated only in the PR400. Quadratic regression analysis showed that the optimal dietary PIP level was estimated as 2.07-2.13 g/kg to maximize growth performance. Overall, these results indicate that dietary PIP at an appropriate level can improve immunity, cytokine gene expression, and disease resistance in C. carpio., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This study evaluated the effects of jamun leaf extract (JLE) as a feed supplement on growth performance, haemato-immunological, oxidative stress-related parameters, and cytokine gene expression in Cyprinus carpio challenged with Aeromonas hydrophila.. Diets containing four different JLE concentrations, that is, 0 (basal diet), 5 (JLE5), 10 (JLE10), and 15 g kg -1 (JLE15), were fed to carp (6.17 ± 0.43 g) for eight weeks. Growth performance was significantly higher in JLE10. Haemato-immunological and antioxidant parameters were determined in fish at 48 h post-challenge with A. hydrohila. The cumulative survival was highest in JLE10 (69.69%) 14 days post-challenge. Serum protein (2.18 ± 0.06 g dL -1 ), lysozyme (32.38 ± 1.2 U mL -1 ), alternative complement pathway (70.43 ± 1.61 U mL -1 ), phagocytic activity (21.18 ± 0.48%), respiratory burst activity (0.289 ± 0.09 OD 630nm ), and immunoglobulin levels (6.67 ± 0.36 U mg mL -1 ) were significantly higher in JLE10 than in the control. Serum alanine aminotransferase (44.06 ± 1.62 Unit mL -1 ), aspartate aminotransferase (31.58 ± 1.82 Unit mL -1 ), and malondialdehyde (2.57 ± 0.26 nmol mL -1 ) levels were lower in JLE10 than in the control (p < 0.05), whereas myeloperoxidase activity was significantly higher in JLE5 and JLE10 than in the control. Superoxide dismutase levels in the serum were higher (p < 0.05) in JLE5 and JLE10 than in the other groups. Gene expression analysis revealed that the mRNA expression of pro-inflammatory cytokines TNF-α and IL-1β was upregulated (p < 0.05) in the liver, head-kidney, and intestine of challenged carp in JLE10. The signalling molecule NF-κB p65 was upregulated in lymphoid organs in JLE10 but not in the liver. The anti-inflammatory cytokine IL-10 was significantly downregulated in challenged carp in JLE10 compared with that in the control. Quadratic regression analysis showed that optimal dietary JLE was estimated to be 9.03-10.15 g kg -1 to maximize the growth performance. Results of the present study revealed that dietary JLE at 10 g kg -1 can significantly improve the immunity and disease resistance of C. carpio. Thus, JLE is a promising food additive for carp aquaculture., Competing Interests: Declaration of competing interest Authors do not have any competing interest to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The recent outbreak of blight in pome fruit plants has been a major concern as there are two indistinguishable Erwinia species, Erwinia amylovora and E. pyrifoliae , which cause blight in South Korea. Although there is a strict management protocol consisting of antibiotic-based prevention, the area and the number of cases of outbreaks have increased. In this study, we isolated four bacteriophages, pEp&#95;SNUABM&#95;03, 04, 11, and 12, that infect both E. amylovora and E. pyrifoliae and evaluated their potential as antimicrobial agents for administration against Erwinia -originated blight in South Korea. Morphological analysis revealed that all phages had podovirus-like capsids. The phage cocktail showed a broad spectrum of infectivity, infecting 98.91% of E. amylovora and 100% of E. pyrifoliae strains. The antibacterial effect was observed after long-term cocktail treatment against E. amylovora , whereas it was observed for both short- and long-term treatments against E. pyrifoliae . Genomic analysis verified that the phages did not encode harmful genes such as antibiotic resistance or virulence genes. All phages were stable under general orchard conditions. Collectively, we provided basic data on the potential of phages as biocontrol agents that target both E. amylovora and E. pyrifoliae.

Long-term feed route exposure to T-2 toxin was proved to elicit growth retarding effects and induction of oxidative stress and apoptosis in Chinese mitten crab (Eriocheir sinensis). However, no study with a holistic perspective has been conducted to date to further describe the in-depth toxicological mechanism of T-2 toxin in E.sinensis. In this study, an RNA-Sequencing (RNA-seq) was used in this study to investigate the effects of feed supplementation with 0 mg/kg and 4 mg/kg T-2 toxin on the hepatopancreas transcriptome of E.sinensis and establish a hepatopancreas transcriptome library of T-2 toxin chronically exposed crabs after five weeks, where 14 differentially expressed genes (DEGs) were screened out across antioxidant, apoptosis, autophagy, glucolipid metabolism and protein synthesis. The actual expression of all the DEGs (Caspase, ATG4, PERK, ACSL, CAT, BIRC2, HADHA, HADHB, ACOX, PFK, eEFe1, eIF4ɑ, RPL13Ae) was also analyzed by real-time quantitative PCR (RT-qPCR). It was demonstrated that long-term intake of large amounts of T-2 toxin could impair antioxidant enzyme activity, promote apoptosis and protective autophagy, disrupt lipid metabolism and inhibit protein synthesis in the hepatopancreas of E.sinensis. In conclusion, this study explored the toxicity mechanism of T-2 toxin on the hepatopancreas of E.sinensis at the mRNA level, which lays the foundation for further investigation of the molecular toxicity mechanism of T-2 toxin in aquatic crustaceans., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

This study evaluated the effects of Bougainvillea glabra (BG) leaf as a feed supplement on growth, skin mucosal immune parameters, serum oxidative stress, expression of immune-related genes, and susceptibility to pathogen infection in carp Cyprinus carpio. Diets containing four different BG concentrations (g kg -1 ), i.e., 0 g (basal diet), 20 g (BG20), 30 g (BG30), 40 g (BG40), and 50 g (BG50), were fed to the carp (average weight: 14.03 ± 0.81 g) for 8 weeks. Skin mucosal immunological and serum antioxidant parameters were examined 8 weeks post-feeding. Growth performance was significantly higher in BG40. Among the examined skin mucosal immune parameters, lysozyme (33.79 ± 0.98 U mL -1 ), protein (6.88 ± 0.37 mg mL -1 ), immunoglobulin (IgM; 5.34 ± 0.37 unit-mg mL -1 ), and protease activity (3.18 ± 0.36%) were significantly higher in BG40 than in the control; whereas, there was no significant effect on the alkaline phosphatase level. Among serum immune activity, activities of lysozyme, the alternative complement pathway, and IgM were significantly higher in BG40. Phagocytic, and superoxide dismutase (SOD) activities were higher (P < 0.05) in BG30-BG50. Serum ALT, AST, and MDA levels were lower in BG40 than in the control (P < 0.05). Intestinal enzymatic activities were enhanced in BG40 and BG50 (P < 0.05), except for lipase in BG50. Gene expression analysis revealed that the mRNA expressions of antioxidant genes (SOD, GPx, and Nrf2), an anti-inflammatory gene (IL-10), and IκBα were significantly upregulated in BG40. Conversely, the pro-inflammatory gene IL-1β and the signaling molecule NF-κB p65 were downregulated in BG40 and BG50, respectively. BG supplementation had no significant effect on TNF-α, TLR22, or HSP70 mRNA expressions. Moreover, fish in BG40 exhibited the highest relative post-challenge survival (67.74%) against Aeromonas hydrophila infection. These results suggested that dietary supplementation with BG leaves at 40 g/kg can significantly improve the growth performance, immune responses, and disease resistance of C. carpio. BG leaves are a promising food additive for carp in aquaculture., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

With the growing demand for caviar products, sturgeon (family Acipenseridae) have become some of the most popular species in the aquaculture industry. Since sturgeon need more than 10 years to become sexually mature, protection against fatal diseases becomes particularly important in the industry. In March 2018, approximately 10% of Siberian Sturgeon Acipenser baerii fingerlings in a sturgeon hatchery in Jeolla Province, Republic of Korea, exhibited anorexia, abdominal distension, buoyancy loss, and abnormal behavior and eventually showed a 90% fatality rate. Twenty moribund fish were necropsied, and a distended stomach filled with gas was found in every case. A single colony dominated the intestinal lumen smear and was identified as Candida manassasensis by polymerase chain reaction targeting 18S ribosomal RNA. The same microorganism was also detected in the sturgeons' feed. Antifungal resistance was examined using the VITEK 2 system, and the isolate was susceptible to voriconazole, caspofungin, micafungin, and flucytosine. The environmental stress factor for this case was speculated as decreased water temperature. Since similar cases have been observed for many years, further research to optimize precise treatment and prevention methods is required., (© 2022 American Fisheries Society.)

Recently, there has been an increasing number of blight disease reports associated with Erwinia amylovora and Erwinia pyrifoliae in South Korea. Current management protocols that have been conducted with antibiotics have faced resistance problems and the outbreak has not decreased. Because of this concern, the present study aimed to provide an alternative method to control the invasive fire blight outbreak in the nation using bacteriophages (phages) in combination with an antibiotic agent (kasugamycin). Among 54 phage isolates, we selected five phages, pEa&#95;SNUABM&#95;27, 31, 32, 47, and 48, based on their bacteriolytic efficacy. Although only phage pEa&#95;SNUABM&#95;27 showed host specificity for E. amylovora , all five phages presented complementary lytic potential that improved the host infectivity coverage of each phage All the phages in the cocktail solution could lyse phage-resistant strains. These strains had a decreased tolerance to the antibiotic kasugamycin, and a synergistic effect of phages and antibiotics was demonstrated both in vitro and on immature wound-infected apples. It is noteworthy that the antibacterial effect of the phage cocktail or phage cocktail-sub-minimal inhibitory concentration (MIC) of kasugamycin was significantly higher than the kasugamycin at the MIC. The selected phages were experimentally stable under environmental factors such as thermal or pH stress. Genomic analysis revealed these are novel Erwinia -infecting phages, and did not encode antibiotic-, virulence-, or lysogenic phage-related genes. In conclusion, we suggest the potential of the phage cocktail and kasugamycin combination as an effective strategy that would minimize the use of antibiotics, which are being excessively used in order to control fire blight pathogens.

With concern growing over antibiotics resistance, the use of bacteriophages to combat resistant bacteria has been suggested as an alternative strategy with which to enable the selective control of targeted pathogens. One major challenge that restrains the therapeutic application of bacteriophages as antibacterial agents is their short lifespan, which limits their antibacterial effect in vivo. Here, we developed a polylactic-co-glycolic acid (PLGA)/alginate-composite microsphere for increasing the lifespan of bacteriophages in vivo. The alginate matrix in PLGA microspheres encapsulated the bacteriophages and protected them against destabilization by an organic solvent. Encapsulated bacteriophages were detected in the tissue for 28 days post-administration, while the bacteriophages administered without advanced encapsulation survived in vivo for only 3-5 days. The bacteriophages with extended fate showed prophylaxis against the bacterial pathogens for 28 days post-administration. This enhanced prophylaxis is presumed to have originated from the diminished immune response against these encapsulated bacteriophages because of their controlled release. Collectively, composite encapsulation has prophylactic potential against bacterial pathogens that threaten food safety and public health.

Natural products have gained considerable attention for improving fish growth performance and immunity to enhance disease resistance. This study evaluated the effect of dandelion polysaccharides (DP) on skin mucosal immune parameters, immune-related gene expression, and susceptibility to pathogen challenge in the Common carp Cyprinus carpio. Diets containing four different concentrations of DP (g Kg -1 ):0 g [basal diet], 0.5 g [D1], 1.5 g [D2], 2.5 g [D3], and 4.0 g [D4] were fed to the carp (average weight: 13.92 ± 0.83 g) for eight weeks. Growth parameters were analyzed four and eight weeks after feeding. Immunological, hematological, and antioxidant parameters were examined eight weeks post-feeding. Growth performance was significantly higher on D3, with a final weight gain of 71.48 ± 1.57 g and a specific growth rate of 3.06 ± 0.12. Among hematological parameters examined, erythrocyte , hematocrit, and mean corpuscular volume (MCV) levels were significantly higher in D3. Skin mucosal immune parameters, such as lysozyme (31.04 ± 1.02 Unit mL -1 ), alkaline phosphatase (122.6 ± 3.8 IU L -1 ), and protein level (10.6 ± 0.74 mg mL -1 ) were significantly higher in D3, while peroxidase activity was higher in D4. Furthermore, SOD activity was higher in D2-D3, whereas catalase activity was higher in D2-D4 (P < 0.05) than in the control. Malondialdehyde level decreased significantly in D3 (5.43 ± 0.36 nmol mL -1 ); whereas, serum ALT and AST levels were significantly lower on D2-D4. Intestinal tight-junction-related genes ZO-1 and Claudin 7 were significantly higher in the DP-fed groups; however, DP had no significant effect on claudin 3. Occludin expression was higher (p < 0.05) on D3 only. Pro-inflammatory cytokines (IL-1β and TNF-α) and IFN-γ strongly upregulated in the head kidney at D3. Conversely, the expression of the anti-inflammatory cytokine interleukin-10, HSP70, and TOR were considerably downregulated in D3. Fish from D3 exhibited markedly higher relative post-challenge survival (66.67%) against Aeromonas hydrophila challenge. The results of the present study suggest that dietary supplements of DP at 2.5 g kg -1 can significantly improve the growth performance, skin mucosal, and serum antioxidant parameters, and strengthen the immunity of C. carpio. Therefore, DP is a promising food additive for carp aquaculture., Competing Interests: Declaration of competing interest Authors do not have any competing interest to declare., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Global warming is favouring the incidence, intensity and duration of harmful cyanobacterial blooms. Microcystin-LR (MC-LR), a hepatotoxic agent, is produced during cyanobacterial blooms. To understand the molecular mechanisms of acute hepatotoxic effect of low doses of MC-LR in crab, we examined differentially expressed genes in samples of the hepatopancreas of Chinese mitten crab (Eriocheir sinensis) collected in 48 h after injections of MC-LR at doses of 0, 25, 50, and 75 µg/kg. The results revealed that MC-LR induced changes in corresponding gene led to the accumulation of triglycerides. MC-LR exposure affected sterol metabolism. Apoptosis-related genes such as Fas-L, Bcl-XL, Cytc, AiF, p53, PERK, calpain, CASP2, CASP7, α-tubulin, PARP, GF, G12, and PKC were upregulated. Conversely, expression levels of CASP10 and ASK1 were downregulated. Genes related to the regulation of actin cytoskeleton (Rho, ROCK, MLCP, MLC, PAK, and PFN) were upregulated. Further, expression levels of genes encoding fatty acid elongation-related enzymes were upregulated, but the expression of genes related to fatty acid synthesis was slightly down regulated. Taken together, these results demonstrated the hepatic toxicity and molecular mechanisms of changes in lipid metabolism, immune and apoptosis in Chinese mitten crab under the MC-LR-induced stress, which is the first report on crabs and performs a comprehensive analysis and a new insight of the molecular toxicological responses in crabs., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonellosis is a disease of critical concern for public health, and the use of bacteriophages is among the most promising approaches to combating Salmonella. As Salmonella has various serotypes and strains, and bacteriophages are virulent to specific hosts, it is important to isolate phages and evaluate interactions with their hosts. In the present study, a novel Salmonella-infecting bacteriophage, pSal-SNUABM-01, was isolated and characterized. Transmission electron microscopy revealed that the bacteriophage is a member of the family Podoviridae and possesses an elongated head and a short tail. The phage genome is circular and 89,500 bp in size. A total of 162 open reading frames were predicted, eight of which were tRNAs. Morphological and genomic analysis revealed that pSal-SNUABM-01 is closely related to phage 7-11. In phylogenetic analysis, pSal-SNUABM-01 and 7-11 did not cluster together with the members of any established genus, suggesting that these two phages comprise a novel genus. The results of this study enhance our understanding of the phylogeny of the family Podoviridae and might be applicable to the development of bacteriophage treatments against Salmonella infections., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

In the aquaculture industry, an efficient and safe water purification system is important to prevent mass mortality by virulent pathogens. As extensive use of traditional methods (e.g.: povidone-iodine, ozone, ultraviolet irradiation, formalin, and chlorine dioxide) have adverse effects on cultured fish, an appropriate and alternative water purification method is vital for the sustainability of the industry. Non-thermal plasma technology has been successfully used for various biomedical purposes (e.g: food sterilization, medical device disinfection, wound healing, cancer therapy, etc.) and has great potential to be used as a sterilizing system. However, few studies have been conducted on its usefulness in the aquaculture industry. In this study, we investigated the bactericidal efficacy of plasma-activated water induced by non-thermal plasma and its histopathological as well as immunological adverse effects on koi. A highly virulent Aeromonas hydrophila SNU HS7, which caused massive mortality of koi, was used for this study. Non-thermal plasma was applied for 10 min to the fish tanks with 1.2 × 10 9  CFU/mL SNU HS7 using PLMB-20 system to confirm the sterilization efficacy and to observe the survival and immunological reaction of koi for 14 days. As a result, gross pathological, histopathological, and immunological investigations did not reveal any significant adverse effects in fish as compared to the control groups. To the best of our knowledge, this is the first study showing that non-thermal plasma can be used for sterilization of rearing water without giving significant physiological damage to the fish, even under the assumption of extreme situations. As plasma can effectively sterilize not only bacteria but also other unknown pathogens, the results of this study are showing a promising future in purifying water in aquaculture practice., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The present study investigated the effect of Pandanus tectorius (PT) extract on Cyprinus carpio growth, antioxidant capacity, immunological and oxidative stress, immune-related gene expression, and resistance against pathogen challenge. Fish (average weight: 12.18 ± 0.32 g) were fed with diets containing various concentration (g kg -1 ) of PT extract: 0 g (basal diet), 5 g [PT5], 10 g [PT10], 20 g [PT20], and 30 g [PT30] for 8 weeks. Our results revealed an increase (p < 0.05) in the final weight gain only in the PT20 (63.12 ± 1.4 g) group. Activity of liver antioxidant parameters such as catalase (CAT) (27.67 ± 1.12 U mg protein -1 ), superoxide dismutase (SOD) (13.17 ± 0.428 U mg protein -1 ), and glutathione peroxidase (GPx) (47.14 ± 1.47 U mg protein -1 ) was highest in the PT20 group. Among the serum immunological parameters examined, lysozyme (37.45 ± 0.67 U mL -1 ), alternative complement pathway (133.22 ± 1.92 U mL -1 ), and phagocytic activities (25.68 ± 0.93%) were high (p < 0.05) in PT20, whereas there was no significant effect on serum immunoglobulin or total protein levels, compared to the control. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and malondialdehyde (MDA) levels were found to be lower (p < 0.05) in the PT20 and PT30 groups than those in the control. However, myeloperoxidase (MPO) levels were significantly higher (p < 0.05) in the PT20 and PT30 groups. Gene expression analysis revealed that mRNA expressions of antioxidant genes (SOD, CAT, nuclear factor erythroid 2-related factor 2) and anti-inflammatory cytokine (IL-10) genes were significantly upregulated in the PT10 - PT30 groups, except for the SOD gene in PT10. Conversely, the signalling molecule NF-κBp65 was downregulated in PT20 and PT30. Expression of Toll-like receptor 22 (TLR22) and hepcidin was significantly upregulated in PT20 and PT30, respectively. Fish in the PT20 group exhibited highest relative post-challenge survival (70.37%) against Aeromonas hydrophila challenge. The results of the present study suggest that dietary supplementation of P. tectorius extract at 20 g kg -1 can significantly improve weight gain, serum antioxidant parameters, strengthen immunity, and increase the disease resistance of C. carpio. Therefore, P. tectorius extract could be exploited for its use as a food additive in aquaculture., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Skin mucosal lymphoid tissues of fish are the first line of defence against pathogen invasion. We investigated the effects of Lactiplantibacillus plantarum subsp. plantarum L7, singularly or in combination with Limosilactobacillus reuteri P16, on mucosal immunity and diseases resistance of carp Cyprinus carpio. C. carpio (average weight: 26.28 ± 1.02 g) were divided into five experimental groups. Fish in each group were fed with one of the following potential probiotic-supplemented diets: control (0 - basal diet), D1 (10 7  CFU/g L7), D2 (10 8  CFU/g L7), D3 (10 9  CFU/g L7), and D4 (10 8  CFU/g L7 + 10 8  CFU/g P16). Eight weeks post-feeding, growth performance was higher in D4, with a final weight gain of 67.18 ± 1.47 g. Results showed a significantly higher skin mucosal lysozyme, alkaline phosphatase, mucus protein level, superoxide dismutase, and catalase activities in D2 and D4 compared to the control. However, potential probiotics had no significant effect on skin mucosal immunoglobulin level. Skin mucus of D4 exhibited stronger inhibition zones against pathogenic bacterial strains. Moreover, digestive enzyme activities (protease, lipase) were highest in D4. Intesinal lactic acid bacterial counts of fish fed combind probiotics (i.e. D4) was significantly higher than the control. Further, supplementation of potential probiotics altered the expression of IL-1β, TNF-α, and IL-10 cytokines. Fish from D4 exhibited significantly higher relative post-challenge survival (69.7%) against Aeromonas hydrophila, followed by D2 (66.67%). Therefore, the inclusion of L. plantarum subsp. plantarum L7 at 10 8  CFU/g or in combination with L. reuteri P16 could enhance the growth performance, mucosal immune responses, and disease resistance of C. carpio., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The bacterial genus Pseudomonas is a common causative agent of infections in veterinary medicine. In this study, we focused on Pseudomonas aeruginosa canine otitis externa isolates. Due to prolonged antibiotic treatment of otitis externa, antibiotic resistance is common and has become a major complication. Many alternatives to antibiotics have been studied, with bacteriophages emerging as the most promising alternatives. Here, we isolated and characterized a novel phage, pPa&#95;SNUABM&#95;DT01, by investigating its morphology, growth, lysis kinetics, and genomic characteristics. Phages have a vigorous capacity to eliminate bacterial cells through bacterial lysis. This capacity is dependent on the multiplicity of infection (MOI), but even at low MOIs, the phage successfully inhibited bacterial regrowth. The phage genome was 265,520 bp in size and comprised 312 putative open reading frames (ORFs). Comparative genome analysis demonstrated that the phage is a novel species in Myoviridae . The nucleotide similarity was moderately high compared with the Pseudomonas virus, Noxifer. However, a phylogenetic analysis and a dot plot indicated that pPa&#95;SNUABM&#95;DT01 is not closely related to the Phikzvirus or Noxifervirus genus but, instead, belongs to a novel one. The genome comparisons also indicate that the phage, pPa&#95;SNUABM&#95;DT01, could be a novel genus.

Curcumin exhibited numerous key activities including antioxidant, anti-inflammatory, and immunostimulatory effects in fish. This study evaluated the protective effects of curcumin (CUR) against lead (Pb)-induced toxicities in fish. Healthy Cyprinus carpio L. individuals were segregated into control, Pb only, Pb+CUR, and CUR only groups. Pb groups were exposed to 1 mg L -1 of Pb, and CUR groups were fed a basal diet supplemented with 15 g kg -1 of CUR. After eight weeks, growth performance, Pb accumulation in tissues, various haemato-biochemical parameters, immune responses, and cytokine gene expression were measured. Dietary CUR effectively decreased Pb accumulation in tissues and increased the survival of Pb-exposed fish. Co-treatment with Pb and CUR reversed alterations in haemato-biochemical parameters, ameliorated Pb-induced oxidative stress, enhanced immune responses, and restored intestinal enzymatic activities. Dietary CUR reversed changes in intestinal microbiota in Pb-exposed fish. Pb-induced upregulation of NF-κBp65 and HSP70 was inhibited by dietary CUR. CUR supplementation upregulated the mRNA levels of SOD, Nrf2, IL-10, and CYP450 1A and attenuated Pb-induced degradation of I κB-α mRNA levels. Overall, CUR antagonizes Pb-induced negative impacts in fish. Thus, dietary CUR had several beneficial effects on immune responses, decreased Pb accumulation in tissues, and reversed Pb-induced oxidative stress in fish. Therefore, CUR plays a protective role in Pb-induced immune toxicity in fish, and, as such, may be suitable as an aqua feed additive for use in carp aquaculture., (Copyright © 2021. Published by Elsevier Inc.)

Species belonging to the genus Erwinia are predominantly plant pathogens. A number of bacteriophages capable of infecting Erwinia have been used for the control of plant diseases such as fire blight. Public repositories provide the complete genome information for such phages, which includes genomes ranging from 30 kb to 350 kb in size. However, limited information is available regarding bacteriophages belonging to the family Siphoviridae . A novel lytic siphophage, pEp&#95;SNUABM&#95;08, which specifically infects Erwinia pyrifoliae , was isolated from the soil of an affected apple orchard in South Korea. A comprehensive genome analysis was performed using the Erwinia -infecting siphophage. The whole genome of pEp&#95;SNUABM&#95;08 comprised 62,784 bp (GC content, 57.24%) with 79 open reading frames. The genomic characteristics confirmed that pEp&#95;SNUABM&#95;08 is a singleton lytic bacteriophage belonging to the family Siphoviridae , and no closely related phages have been reported thus far. Our study not only characterized a unique phage, but also provides insight into the genetic diversity of Erwinia bacteriophages.

Copper shark ( Carcharhinus brachyurus Günther, 1870) is one of the most widely distributed but least known species in the family Carcharhinidae. Herein, we report the first complete mitogenome of C. brachyurus . The overall structure of the 16,704 bp C. brachyurus mitogenome was similar to that of other Carcharhinus species and showed the highest average nucleotide identity (97.1%) with the spinner shark ( Carcharhinus brevipinna ). Multigene phylogeny using 13 protein-coding genes (PCGs) in the mitogenome resolved C. brachyurus clustered with other species within the genus; the overall tree topology was congruent with recent phylogenetic studies of this species. These results provide important information for conservation genetics and further evolutionary studies of sharks., Competing Interests: No potential conflict of interest was reported by the authors., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

As a primary bacterial pathogen in companion animals, Staphylococcus pseudintermedius has zoonotic potential. This pathogen exhibits multidrug resistance, including resistance to methicillin, and biofilm-forming ability, making it hard to eradicate with antimicrobial agents. One potential alternative is bacteriophage therapy. In this study, we first characterized the antimicrobial resistance profile of S. pseudintermedius from canine samples and isolated two novel bacteriophages, pSp-J and pSp-S, from canine pet parks in South Korea to potentially control S. pseudintermedius . The biological characteristics of phages were assessed, and the phages could infect most of the methicillin-resistant S. pseudintermedius strains. We found that these phages were stable under the typical environment of the body (~37°C, pH 7). We also assessed bacterial lysis kinetics using the two phages and their cocktail, and found that the phages could prevent biofilm formation at low doses and could degrade biofilm at high doses. Taken together, this study demonstrates that bacteriophages pSp-J and pSp-S isolated in this study can be used to potentially treat methicillin-resistant S. pseudintermedius ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kim, Giri, Yun, Kim, Han, Kwon, Oh, Lee, Park and Park.)

Soft-shell turtle (SST; freshwater terrapin or tortoise) is a popular and important health functional food (HFF) product in many Asian countries. HFFs containing SST must be safe, but several HFFs have been found to be contaminated with dangerous substances, such as nitrofuran metabolites (NFMs). This finding suggests that the consumption of HFFs results in the regular exposure of vulnerable individuals to hazardous substances. Importantly, nitrofuran antibiotics have been banned for use in food-producing animals since the 1990s by the European Union. Thus, in this study, we propose a reliable and quick method to reduce the time required for the detection of four NFMs in SST powder that conventional methods are unable to quantify. Our method involves the derivatization and hydrolysis of SST powder and was validated in accordance with the requirements of European Commission Decision 2002/657/EC. The method achieves an apparent mean recovery of 82.2-108.1%, repeatability of 1.5-3.8%, and reproducibility of 2.2-4.8% for 0.5-10.0  μ g kg -1 of 1-aminohydantoin, semicarbazide, 3-amino-2-oxazolidinone, and 3-amino-5-morpholinomethyl-2-oxazolidinone. In addition, linearity was achieved with correlation coefficients of 0.999, and the detection capability (CC β ) and decision limit (CC α ) were found to be reliable, indicating that this is a fast and accurate method for the analysis of SST powder. The validated method was successfully applied to detect NFMs in SST powder in commercial HHFs., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright © 2021 EunChae Ryu et al.)

Background: Antibiotic-resistant bacteria have emerged as a serious problem; bacteriophages have, therefore, been proposed as a therapeutic alternative to antibiotics. Several authorities, such as pharmacopeia, FDA, have confirmed their safety, and some bacteriophages are commercially available worldwide. The demand for bacteriophages is expected to increase exponentially in the future; hence, there is an urgent need to mass-produce bacteriophages economically. Unlike the replication of non-lytic bacteriophages, lytic bacteriophages are replicated by lysing host bacteria, which leads to the termination of phage production; hence, strategies that can prolong the lysis of host bacteria in bacteria-bacteriophage co-cultures, are required., Results: In the current study, we manipulated the inoculum concentrations of Staphylococcus aureus and phage pSa-3 (multiplicity of infection, MOI), and their energy sources to delay the bactericidal effect while optimizing phage production. We examined an increasing range of bacterial inoculum concentration (2 × 10 8 to 2 × 10 9  CFU/mL) to decrease the lag phase, in combination with a decreasing range of phage inoculum (from MOI 0.01 to 0.00000001) to delay the lysis of the host. Bacterial concentration of 2 × 10 8  CFU/mL and phage MOI of 0.0001 showed the maximum final phage production rate (1.68 × 10 10 plaque forming unit (PFU)/mL). With this combination of phage-bacteria inoculum, we selected glycerol, glycine, and calcium as carbon, nitrogen, and divalent ion sources, respectively, for phage production. After optimization using response surface methodology, the final concentration of the lytic Staphylococcus phage was 8.63 × 10 10  ± 9.71 × 10 9  PFU/mL (5.13-fold increase)., Conclusions: Therefore, Staphylococcus phage pSa-3 production can be maximized by increasing the bacterial inoculum and reducing the seeding phage MOI, and this combinatorial strategy could decrease the phage production time. Further, we suggest that response surface methodology has the potential for optimizing the mass production of lytic bacteriophages.

Salmonellosis is a major global public health issue; its most common infection, gastroenteritis, accounts for approximately 90 million illnesses and 150,000 mortalities per year. Eradicating salmonellosis requires surveillance, prevention and treatment, entailing large expenditures. However, it is difficult to control Salmonella transmission because it occurs via multiple routes; exotic reptiles are a reservoir of Salmonella and comprise one such route. As the popularity of exotic pets and animal exhibition has increased, human encounters with reptiles have also increased. As a result, reptile-associated salmonellosis (RAS) has been recognized as an emerging disease. The development of antimicrobial resistance in RAS-causing Salmonella sp. requires alternatives to antibiotics. In this study, bacteriophages have been established as an alternative to antibiotics because only target bacteria are lysed; thus, they are promising biocontrol agents. Here, bacteriophage pSal-SNUABM-02, which infects and lyses reptile Salmonella isolates, was isolated and characterized. The morphology, host range, growth traits and stability of the phage were investigated. The phage was assigned to Myoviridae and was stable in the following conditions: pH 5-9, 4-37°C, and ultravioletA/ultravioletB (UVA/UVB) exposure. Salmonella clearance efficacy was tested using planktonic cell lysis activity and biofilm degradation on polystyrene 96-well plates and reptile skin fragments. The phage exhibited vigorous lysis activity against planktonic cells. In in vitro biofilm degradation tests on reptile skin and polystyrene plates, both low- and high-concentration phage treatments lowered bacterial cell viability by approximately 2.5-3 log colony-forming units and also decreased biomass. Thus, bacteriophages are a promising alternative to antibiotics for the prevention and eradication of RAS., (© 2021 Wiley-VCH GmbH.)

This study presents a novel Janthinobacterium strain, SNU WT3, isolated from the kidney of rainbow trout. A phylogenetic study using 16S rRNA sequences indicated that the strain is closely related to Janthinobacterium svalbardensis JA-1 T . However, biochemical analysis found differences in D-xylose adonitol, N-acetylglucosamine, arbutin, and cellobiose. As for genome-to-genome distance and average nucleotide identity values calculated between strain SNU WT3 and other related strains such as J. lividum EIF1, J. svalbardensis PAMC 27463, and J. agaricidamnosum BHSEK were all below the cutoff value between species. DNA-DNA hybridization between strain SNU WT3 and other close relatives indicated the results of J. lividum DSM 1522 T (47.11%) and J. svalbardensis JA-1 T (38.88%) individually. The major fatty acid compositions of strain SNU WT3 were cylco-C 17:0 (41.45%), C 16:0 (33.86%) and C 12:0 (5.87%). The major polar lipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, and diphosphatidylglycerol. The quinone system was composed mainly of ubiquinone Q-8. The genome of strain SNU WT3 consists of 6,314,370 bp with a G + C content of 62.35%. Here, we describe a novel species of the genus Janthinobacterium , and the name Janthinobacterium tructae has been proposed with SNU WT3 T (=KCTC 72518 = JCM 33613) as the type strain.

A simplified method is described for reducing the analysis time of nitrofurans (NFs) and nitrofuran metabolites (NFMs) in the aquatic animals. Most existing HPLC-MS/MS methods are intended only for NFMs and are based on their fast metabolic transformations. We optimized a method for simultaneously detecting major NFs and their metabolites, including nitrovin (NV) that imply use of an optimized buffer solution. The novel method was validated by six different aquatic animal matrices (loach, catfish, shrimp, lobster, scallop, and eel) spiked with the analytes at 0.5, 1.0, and 2.0 μg kg -1 . Recovery rates and %RSDs (relative standard deviations) of 82-97% and 1-8% were observed for NFMs, respectively, with values of 70-96% and 1-8% obtained for furazolidone, furaltadone, nitrofurazone, nitrofurantoin, and NV, respectively. Linearity was observed in the 0.1-20 μg L -1 range, with correlation coefficients greater than 0.99 recorded for all compounds. The developed method is sensitive, accurate, easier to use, and faster than previous methods when applied to real samples. To the best of our knowledge, this is the first method that can simultaneously determine NFs and their metabolites, as well as NV, using a single-step extraction process.

A novel Citrobacter species was isolated from the kidney of diseased rainbow trout ( Oncorhynchus mykiss ) reared on a trout farm. Biochemical characterization and phylogenetic analysis were performed for bacterial identification. Sequencing of the 16S rRNA gene and five housekeeping genes indicated that the strain belongs to the Citrobacter genus. However, multilocus sequence analysis, a comparison of average nucleotide identity, and genome-to-genome distance values revealed that strain SNU WT2 is distinct and forms a separate clade from other Citrobacter species. Additionally, the phenotype characteristics of the strain differed from those of other Citrobacter species. Quinone analysis indicated that the predominant isoprenoid quinone is Q-10. Furthermore, strain virulence was determined by a rainbow trout challenge trial, and the strain showed resistance to diverse antibiotics including β-lactams, quinolone, and aminoglycosides. The complete genome of strain SNU WT2 is 4,840,504 bp with a DNA G + C content of 51.94% and 106,068-bp plasmid. Genome analysis revealed that the strain carries virulence factors on its chromosome and antibiotic resistance genes on its plasmid. This strain represents a novel species in the genus Citrobacter for which the name C. tructae has been proposed, with SNU WT2 (=KCTC 72517 = JCM 33612) as the type strain.

Due to their important phylogenetic position among extant vertebrates, sharks are an invaluable group in evolutionary developmental biology studies. A thorough understanding of shark anatomy is essential to facilitate these studies and documentation of this iconic taxon. With the increasing availability of cross-sectional imaging techniques, the complicated anatomy of both cartilaginous and soft tissues can be analyzed non-invasively, quickly, and accurately. The aim of this study is to provide a detailed anatomical description of the normal banded houndshark (Triakis scyllium) using computed tomography (CT) and magnetic resonance imaging (MRI) along with cryosection images. Three banded houndsharks were scanned using a 64-detector row spiral CT scanner and a 3 T MRI scanner. All images were digitally stored and assessed using open-source Digital Imaging and Communications in Medicine viewer software in the transverse, sagittal, and dorsal dimensions. The banded houndshark cadavers were then cryosectioned at approximately 1-cm intervals. Corresponding transverse cryosection images were chosen to identify the best anatomical correlations for transverse CT and MRI images. The resulting images provided excellent detail of the major anatomical structures of the banded houndshark. The illustrations in the present study could be considered as a useful reference for interpretation of normal and pathological imaging studies of sharks.

The increasing emergence of antimicrobial resistance has become a global issue. Therefore, many researchers have attempted to develop alternative antibiotics. One promising alternative is bacteriophage. In this study, we focused on a jumbo-phage infecting Salmonella isolated from exotic pet markets. Using a Salmonella strain isolated from reptiles as a host, we isolated and characterized the novel jumbo-bacteriophage pSal-SNUABM-04. This phage was investigated in terms of its morphology, host infectivity, growth and lysis kinetics, and genome. The phage was classified as Myoviridae based on its morphological traits and showed a comparatively wide host range. The lysis efficacy test showed that the phage can inhibit bacterial growth in the planktonic state. Genetic analysis revealed that the phage possesses a 239,626-base pair genome with 280 putative open reading frames, 76 of which have a predicted function and 195 of which have none. By genome comparison with other jumbo phages, the phage was designated as a novel member of Machinavirus composed of Erwnina phages.

Jumbo phages, which have a genome size of more than 200 kb, have recently been reported for the first time. However, limited information is available regarding their characteristics because few jumbo phages have been isolated. Therefore, in this study, we aimed to isolate and characterize other jumbo phages. We performed comparative genomic analysis of three Erwinia phages (pEa&#95;SNUABM&#95;12, pEa&#95;SNUABM&#95;47, and pEa&#95;SNUABM&#95;50), each of which had a genome size of approximately 360 kb (32.5% GC content). These phages were predicted to harbor 546, 540, and 540 open reading frames with 32, 34, and 35 tRNAs, respectively. Almost all of the genes in these phages could not be functionally annotated but showed high sequence similarity with genes encoded in Serratia phage BF, a member of Eneladusvirus . The detailed comparative and phylogenetic analyses presented in this study contribute to our understanding of the diversity and evolution of Erwinia phage and the genus Eneladusvirus .

Vibrio coralliilyticus ( V. coralliilyticus ) is a pathogen that causes mass mortality in marine bivalve hatcheries worldwide. In this study, we used a bacteriophage (phage) cocktail to prevent multiple-antibiotic-resistant (MAR) and phage-resistant (PR) V. coralliilyticus infection in Pacific oyster ( Crassostrea gigas ) larvae. To prevent the occurrence of phage-resistant strains and decrease the effect of mono-phage treatment, we prepared a phage cocktail containing three types of V. coralliilyticus -specific phages and tested its prophylactic efficacy against MAR and PR V. coralliilyticus infection. The results of the cell lysis test showed that the phage cocktail showed an excellent bactericidal effect against the MAR and PR variants in contrast to the experimental group treated with two mono phages (pVco-5 and pVco-7). An in vivo test using Pacific oyster larvae also confirmed the preventive effect against MAR and PR variants. In conclusion, the application of the phage cocktail effectively prevented V. coralliilyticus infection in marine bivalve seedling production. Furthermore, it is expected to reduce damage to the aquaculture industry caused by the occurrence of MAR and PR V. coralliilyticus . Therefore, phage cocktails may be used for the control of various bacterial diseases., Competing Interests: The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

The anti-inflammatory activity of the guava leaf extracts (GLE) against LPS-induced inflammatory responses in fish macrophage cell lines is well documented. Here, we evaluated the effects of dietary GLE on LPS-induced oxidative stress, immune responses, and glucocorticoid receptor-related gene expression in Cyprinus carpio. Basal diet was supplemented with 0 (control), 100, 150, 200, or 250 mg kg -1  GLE for eight weeks. Highest (p < 0.05) weight gain rate was obtained in fish group supplemented with 200 mg kg -1 of GLE. The results showed that superoxide dismutase, glutathione-S-transferase, glutathione peroxidase, glutathione reductase, lysozyme, and complement C3 decreased, while malondialdehyde level increased in the liver and spleen upon LPS-challenge. Dietary GLE supplementation (especially 200 or 250 mg kg -1 ) alleviated LPS-induced changes. Similarly, GLE (150-250 mg kg -1 ) reversed LPS-induced alteration of serum biochemical parameters such as alkaline phosphatase, aspartate transaminase, alanine transaminase, and myeloperoxidase. LPS treatment markedly induced increased the mRNA levels of TNF-α, IL-1β, and NF-κB p65 in both the liver and kidney tissues; however, GLE pre-treatment attenuated LPS-induced elicitation of TNF-α, IL-β, and NF-κB p65. Moreover, dietary GLE supplementation significantly increased the expression of HSP70 and HSP90, and glucocorticoid receptor in the liver and kidney after LPS challenge. Thus, GLE attenuated LPS-induced inflammation response by up-regulating glucocorticoid receptor-related gene expression in carp. Finally, GLE supplementation reduced carp mortality after LPS-challenge. These results suggest that dietary supplementation with 200 mg kg -1  GLE is adequate for effectively attenuating LPS-induced oxidative stress and immune-suppressive effects in C. carpio., (Copyright © 2020. Published by Elsevier Ltd.)

Microbial surfactants (biosurfactants) are a broad category of surface-active biomolecules with multifunctional properties. They self-assemble in aqueous solutions and are adsorbed on various interfaces, causing a decrease in surface tension, as well as interfacial tension, solubilization of hydrophobic compounds, and low critical micellization concentrations. Microbial biosurfactants have been investigated and applied in several fields, including bioremediation, biodegradation, food industry, and cosmetics. Biosurfactants also exhibit anti-microbial, anti-biofilm, anti-cancer, anti-inflammatory, wound healing, and immunomodulatory activities. Recently, it has been reported that biosurfactants can increase the immune responses and disease resistance of fish. Among various microbial surfactants, lipopeptides, glycolipids, and phospholipids are predominantly investigated. This review presents the various immunological activities of biosurfactants, mainly glycolipids and lipopeptides. The applications of biosurfactants in aquaculture, as well as their immunomodulatory activities, that make them novel therapeutic candidates have been also discussed in this review.

Currently, topical antibiotic treatment is a major strategy for decolonisation of Staphylococcus aureus, although it may result in antibiotic resistance or recolonisation of the organism. Recently, application of bacteriophages in the treatment of S. aureus infection has attracted attention. However, a single administration of bacteriophages did not effectively decolonise S. aureus in our first trial in vivo. Using a bacteriophage (pSa-3) and surfactant combination in vitro, we showed an increased (>8%) adsorption rate of the bacteriophage on the host. Moreover, the combination increased the eradication of immunoglobulin E (IgE)-stimulated aggregation, as the surfactant promoted the dissociation of S. aureus aggregates by decreasing the size by 75% and 50% in the absence and presence of IgE, respectively. Furthermore, the combined treatment significantly decolonised the pathogen with an efficacy double that of the phage-only treatment, and decreased the expression of pro-inflammatory cytokine genes (IL-1β, IL-12 and IFNγ) for 5 days in the second in vivo trial. These results suggest that the bacteriophage-surfactant combination could act as an alternative to antibiotics for S. aureus decolonisation in patients with dermatitis., (Copyright © 2020 Elsevier Ltd. All rights reserved.)