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Molecular Immunology Sep 2021As a proinflammatory cytokine of the interleukin-1 (IL-1) family, IL-18 plays important roles in host protection against bacterial, viral, and fungal infection. We...
As a proinflammatory cytokine of the interleukin-1 (IL-1) family, IL-18 plays important roles in host protection against bacterial, viral, and fungal infection. We cloned the open reading frame of snakehead (Channa argus) IL-18 (shIL-18) and found that it contained 609 base pairs and encoded 202 amino acid residues. The shIL-18 included a conserved IL-1-like family signature and two potential IL-1β-converting enzyme cutting sites; one was conserved in all analyzed IL-18s, but the other was unique to shIL-18. Unlike other IL-18s, shIL-18 also contained a predicted signal peptide. In this study, shIL-18 was constitutively expressed in all tested tissues, and its expression was induced by Aeromonas schubertii and Nocardia seriolae in the head kidney and spleen in vivo and by lipoteichoic acid, lipopolysaccharides, and polyinosinic-polycytidylic acid in head kidney leukocytes in vitro. Moreover, recombinant shIL-18 upregulated the expression of interferon-γ, IL-1β, and tumor necrosis factor-α1 and -α2 and promoted the proliferation of leukocytes. Taken together, these results showed that IL-18 played crucial roles in host defense against bacterial infection in fish, as it does in mammals.
Topics: Aeromonas; Animals; Cloning, Molecular; Fish Diseases; Fish Proteins; Fishes; Gram-Negative Bacterial Infections; Head Kidney; Interleukin-18; Lipopolysaccharides; Nocardia; Nocardia Infections; Spleen; Teichoic Acids
PubMed: 34280771
DOI: 10.1016/j.molimm.2021.07.013 -
Comparative Biochemistry and... 2021Mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 3 (MEKK3) is a serine/threonine protein kinase that acts as a key regulator and is...
MEKK3 in hybrid snakehead (Channa maculate ♀ ×Channa argus ♂): Molecular characterization and immune response to infection with Nocardia seriolae and Aeromonas schubertii.
Mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase 3 (MEKK3) is a serine/threonine protein kinase that acts as a key regulator and is widely involved in various innate and acquired immune signaling pathways. In this study, we first cloned the complete open reading frame (ORF) of the MEKK3 gene (named CcMEKK3) in a hybrid snakehead (Channa maculate ♀ × Channa argus ♂). The full-length ORF of CcMEKK3 is 1851 bp, and encodes a putative protein of 616 amino acids containing a serine/threonine kinase catalytic (S-TKc) domain and a Phox and Bem1p (PB1) domain. A sequence alignment and phylogenetic tree analysis showed that CcMEKK3 is highly conserved relative to the MEKK3 proteins of other teleost species. CcMEKK3 was constitutively expressed in all the healthy hybrid snakehead tissues tested, with greatest expression in the immune tissues, such as the head kidney and spleen. The expression of CcMEKK3 was usually upregulated in the head kidney, spleen, and liver at different time points after infection with Nocardia seriolae or Aeromonas schubertii. Similarly, the dynamic expression levels of CcMEKK3 in head kidney leukocytes after stimulation revealed that CcMEKK3 was induced by LTA, LPS, and poly(I:C). In the subcellular localization analysis, CcMEKK3 was evenly distributed in the cytoplasm of HEK293T cells, and its overexpression significantly promoted the activities of NF-κB and AP-1. These results suggest that CcMEKK3 is involved in the immune defense against these two pathogens, and plays a crucial role in activating the NF-κB and MAPK signaling pathways.
Topics: Aeromonas; Animals; Fish Diseases; Fish Proteins; Fishes; Gram-Negative Bacterial Infections; Immunity, Innate; MAP Kinase Kinase Kinase 3; Nocardia; Nocardia Infections
PubMed: 34186154
DOI: 10.1016/j.cbpb.2021.110643 -
Microbial Pathogenesis Dec 2020Retrospective diagnosis of a bacterial collection (n = 31) originated from five farms reportedly affected by early mortality syndrome (EMS) in Southeast Asia in 2016...
Retrospective diagnosis of a bacterial collection (n = 31) originated from five farms reportedly affected by early mortality syndrome (EMS) in Southeast Asia in 2016 revealed that 9/31 isolates from two farms tested positive for V. parahaemolyticus causing acute hepatopancreatic necrosis disease (VP). Molecular analysis of the 22 remaining isolates showed that 21 isolates belong to Vibrio species including VP, V. vulnificus, V. cholerae, V. owensii and V. alginolyticus. One isolate from an AHPND farm was preliminarily identified as Aeromonas schubertii based on 99.43% nucleotide identity of 16S rRNA to the reference strain ATCC 43700 (X60416). Diseases caused by Vibrio bacteria have been well-studied in shrimp while pathogenic potential of non-Vibrio species has been relatively overlooked. Since the description of A. schubertii present in shrimp farms is rare, this study therefore focused on species identification and its pathogenic potential to shrimp based on a combination of multiple approaches i.e. multilocus sequence analysis (MLSA), challenge test, histopathology and in situ hybridization (ISH). Based on MLSA of 2464 bp derived from 16S rRNA (1346 bp), gyrB (568 bp) and rpoB (550 bp), this isolate was confirmed as A. schubertii. Immersion challenge using three successive 10-fold serial dilutions (2 × 10 to 2 × 10 CFU/mL) revealed that A. schubertii was pathogenic to shrimp and cumulative mortalities were dose-dependent (45-70%). The diseased shrimp exhibited gross sign of reddish body and remarkable histopathological lesion of collapsed hepatopancreatic tubules and typical encapsulation. ISH using A. schubertii-specific probe confirmed localization of bacteria in the hepatopancreas of the infected shrimp. In summary, this study reported a novel pathogenic, non-Vibrio species, A. schubertii recovered from an AHPND-affected farm causing up to 70% mortality in immersion challenge. Since A. schubertii is relatively new to shrimp, this may pose a potential risk for low salinity shrimp farming areas, active surveillance of this pathogen, therefore, should not be overlooked.
Topics: Aeromonas; Animals; Penaeidae; RNA, Ribosomal, 16S; Retrospective Studies; Vibrio; Vibrio parahaemolyticus
PubMed: 32950638
DOI: 10.1016/j.micpath.2020.104501 -
Fish & Shellfish Immunology Sep 2020Tumor necrosis factor-α (TNF-α) is a pluripotent mediator of pro-inflammatory and antimicrobial defense mechanisms and a regulator of lymphoid organ development....
Tumor necrosis factor-α (TNF-α) is a pluripotent mediator of pro-inflammatory and antimicrobial defense mechanisms and a regulator of lymphoid organ development. Although two types of TNF-α have been identified in several teleost species, their functions in pathogen infection remain largely unexplored, especially in pathogen clearance. Herein, we cloned and characterized two types of TNF-α, termed shTNF-α1 and shTNF-α2, and their receptors, shTNFR1 and shTNFR2, from snakehead (Channa argus). These genes were constitutively expressed in all tested tissues, and were induced by Aeromonas schubertii and Nocardia seriolae in head kidney and spleen in vivo, and by lipoteichoic acid (LTA), lipopolysaccharides (LPS), and Polyinosinic-polycytidylic acid [Poly (I:C)] in head kidney leukocytes (HKLs) in vitro. Moreover, recombinant shTNF-α1 and shTNF-α2 upregulated the expression of endogenous shTNF-α1, shTNF-α2, shTNFR1, and shTNFR2, and enhanced intracellular bactericidal activity, with shTNF-α1 having a greater effect than shTNF-α2. These findings suggest important roles of fish TNFα1, TNFα2, and their receptors in bacterial infection and pathogen clearance, and provide a new insight into their function in antibacterial innate immunity.
Topics: Aeromonas; Animals; Fish Diseases; Fish Proteins; Fishes; Gram-Negative Bacterial Infections; Head Kidney; Immunity, Innate; Leukocytes; Lipopolysaccharides; Nocardia; Nocardia Infections; Poly I-C; Receptors, Tumor Necrosis Factor; Spleen; Teichoic Acids; Tumor Necrosis Factor-alpha
PubMed: 32585357
DOI: 10.1016/j.fsi.2020.05.059 -
Fish & Shellfish Immunology May 2020As a central pro-inflammatory cytokine, interleukin-1β (IL-1β) plays critical roles in the inflammatory response, pathogen infection, and immunological challenges in...
As a central pro-inflammatory cytokine, interleukin-1β (IL-1β) plays critical roles in the inflammatory response, pathogen infection, and immunological challenges in mammals. Although fish IL-1β has been confirmed to participate in inflammatory response to pathogen infection, few studies have been performed to characterize the antibacterial and bactericidal functions of fish IL-1β. In this study, snakehead (Channa argus) IL-1β (shIL-1β) and its receptors, shIL-1R1 and shIL-1R2, were cloned and functionally characterized. ShIL-1β contained the IL-1 family signature domain, and a potential cutting site at Asp that presented in all vertebrate IL-1β sequences. ShIL-1R1 had three extracellular IG-like domains and one intracellular signal TIR domain, while shIL-1R2 had three extracellular IG-like domain but lacked the intracellular signal TIR domain. ShIL-1β, shIL-1R1, and shIL-1R2 were constitutively expressed in all tested tissues, and their expressions could be induced by Aeromonas schubertii and Nocardia seriolae in the head kidney and spleen in vivo, and by LTA, LPS, and Poly (I:C) in head kidney leukocytes (HKLs) in vitro. Moreover, recombinant shIL-1β upregulated the expression of endogenous shIL-1β, shIL-R1, and shIL-R2 in snakehead HKLs, and enhanced intracellular bactericidal activity. Taken together, this study found that, like IL-1β and its receptors in mammals, shIL-1β and its receptors play crucial roles in antibacterial innate immunity. This provides new insight into the evolution of IL-1β function in vertebrates.
Topics: Animals; Anti-Bacterial Agents; Bacteria; Bacterial Infections; Carps; Cloning, Molecular; Fish Diseases; Head Kidney; Immunity, Innate; Interleukin-1beta; Receptors, Interleukin-1
PubMed: 32173451
DOI: 10.1016/j.fsi.2020.03.022 -
Applied Microbiology and Biotechnology Nov 2019Streptococcus agalactiae is a major pathogen causing streptococcosis. To prevent and control this bacterial disease, antagonistic bacteria have become a new research...
Bacillus velezensis LF01: in vitro antimicrobial activity against fish pathogens, growth performance enhancement, and disease resistance against streptococcosis in Nile tilapia (Oreochromis niloticus).
Streptococcus agalactiae is a major pathogen causing streptococcosis. To prevent and control this bacterial disease, antagonistic bacteria have become a new research hotspot. This study evaluated the probiotic potential of Bacillus velezensis LF01 strain, which is antagonistic to S. agalactiae. The active compounds produced by LF01 showed antimicrobial activity against a broad spectrum of fish pathogens, including S. agalactiae, Streptococcus iniae, Aeromonas hydrophila, Edwardsiella tarda, Edwardsiella ictaluri, Aeromonas schubertii, Aeromonas veronii, Aeromonas jandaei, and Vibrio harveyi. The antimicrobial compounds were heat stable, pH stable, UV stable, resistant to proteases, and could be stored for a long time. To evaluate the probiotic function of LF01 in Nile tilapia, juveniles were divided into three treatment groups: a control group, an interval feeding group, and a continuous feeding group. Tilapia fed with LF01-supplemented diets (1.0 × 10 CFU/g) showed significantly better growth performances than those of the control group (P < 0.05). Tilapia fed with LF01-supplemented diets significantly increased lysozyme (LZY) and superoxide dismutase (SOD) activities. The expression of three immune-related genes (C3, lyzc, and MHC-IIβ) was higher in the intestine, head kidney, and gill of tilapia from the continuous feeding group than in those from the control group (P < 0.05). Tilapia fed with LF01-supplemented diets showed remarkably improved survival rates after S. agalactiae infection, and analysis of their intestinal tract pathogens revealed that the abundance of Edwardsiella and Plesiomonas had significantly decreased compared with the control group. Our findings demonstrate that LF01 is an effective antagonist against various fish pathogens and has potential for controlling infections by Streptococcus spp. and other pathogens in tilapia.
Topics: Animals; Antibiosis; Bacillus; Biological Control Agents; Cichlids; Fish Diseases; High-Throughput Nucleotide Sequencing; Probiotics; Streptococcal Infections; Streptococcus agalactiae
PubMed: 31654082
DOI: 10.1007/s00253-019-10176-8