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Animals : An Open Access Journal From... Mar 2024is a pathogen that severely affects aquatic animals, including the snakehead, . Lytic bacteriophages have been recognized as effective alternatives to antibiotics for...
is a pathogen that severely affects aquatic animals, including the snakehead, . Lytic bacteriophages have been recognized as effective alternatives to antibiotics for controlling bacterial infections. However, there have been no reports of phages as far as we know. In this study, a lytic bacteriophage SD04, which could effectively infect , was isolated from pond water cultured with diseased snakehead. The SD04 phage formed small, round plaques on Petri dishes. Electron microscopy revealed a hexagonal head and a contractile tail. Based on its morphology, it may belong to the Myoviridae family. Two major protein bands with molecular weights of 50 and 38 kilodaltons were observed after the phage was subjected to SDS-PAGE. The phage showed a large average burst size, high specificity, and a broad host range. When stored at 4 °C, phage SD04 had high stability over 12 months and showed almost no variation within the first six months. All fish were healthy after both intraperitoneal injection and immersion administration of SD04, indicating the safety of the phage. After treatment with SD04, in both phage therapy groups and prevention groups showed high survival rates (i.e., 83.3 ± 3.3% and 100 ± 1.3%, respectively). Phage therapy inhibits bacterial growth in the liver, the target organ of the infected . The experimental results indicate the potential use of phage SD04 for preventing infection in .
PubMed: 38540055
DOI: 10.3390/ani14060957 -
Journal of Fish Diseases Apr 2024The purpose of this study was to investigate whether a defensin-like antimicrobial peptide (C-13326 peptide) identified in Hermetia illucens could possess protective...
A novel defensin-like peptide C-13326 possesses protective effect against multidrug-resistant Aeromonas schubertii in hybrid snakehead (Channa maculate ♀ × Channa argus ♂).
The purpose of this study was to investigate whether a defensin-like antimicrobial peptide (C-13326 peptide) identified in Hermetia illucens could possess protective effect against multidrug-resistant Aeromonas schubertii in hybrid snakehead (Channa maculate ♀ × Channa argus ♂). The cDNA of C-13326 peptide comprised 243 nucleotides encoding 80 amino acids, with six conserved cysteine residues and the classical CSαβ structure. The recombinant expression plasmid pPIC9K-C-13326 was constructed and transformed into GS115 Pichia pastoris, and the C-13326 peptide was expressed by induction with 1% methanol. The crude extract of C-13326 peptide was precipitated by ammonium sulfate, assayed by Braford method, detected by tricine-SDS-PAGE, evaluated by BandScan software and identified by liquid chromatography-mass spectrometry. The C-13326 peptide was shown to have inhibitory activity against the growth of multidrug-resistant A. schubertii DM210910 by using the minimum growth inhibitory concentration and Oxford cup method. In addition, scanning electron microscopy analysis suggested that C-13326 peptide inhibited the growth of A. schubertii DM210910 by damaging the bacterial cell membrane. To explore the role of peptide C-13326 in vivo, hybrid snakehead was fed with peptide C-13326 as feed additives for 7 days. The results revealed that C-13326 peptide could significantly down-regulate the expression levels of IL-1β, IL-8, IL-12 and TNF-α (p < .05), and significantly improved the survival rate of hybrid snakehead after challenging with A. schubertii DM210910. Therefore, the C-13326 peptide is a promising antimicrobial agent for A. schubertii treatment in aquaculture.
Topics: Animals; Fish Diseases; Fishes; Aeromonas; Peptides; Defensins
PubMed: 38204197
DOI: 10.1111/jfd.13922 -
Frontiers in Immunology 2022β-glucan is widely used in aquaculture due to its immunostimulatory effects, but the specific effect and potential regulatory mechanism on largemouth bass () are still...
β-glucan is widely used in aquaculture due to its immunostimulatory effects, but the specific effect and potential regulatory mechanism on largemouth bass () are still unclear. Here, we evaluated the effects of β-glucan on growth, resistance to , intestinal health, and transcriptome of largemouth bass to reveal the potential regulators, metabolic pathways, and altered differential microbiota. Four experimental diets were designed with β-glucan supplementation levels of 0 (control), 100 (LA-100), 200 (MA-200), and 300 (HA-300) mg kg, and each diet was fed to largemouth bass (79.30 ± 0.50 g) in triplicate for 70 days, followed by a 3-day challenge experiment. Results showed that different β-glucan supplementations had no significant effects on growth performance and whole-body composition. Fish fed a diet with 300 mg kg β-glucan significantly increased the activity of lysozyme than those fed diets with 0 and 100 mg kg β-glucan. In addition, the survival rate of largemouth bass in β-glucan supplementation groups was significantly higher than the control group at 12- and 24-h challenge by . Transcriptome analysis showed that a total of 1,245 genes were differentially expressed [|log(fold change)| ≥1, -value ≤0.05], including 109 immune-related differentially expressed genes (DEGs). Further analysis revealed that significantly upregulated and downregulated DEGs associated with immunity were mapped into 12 and 24 pathways, respectively. Results of intestinal microflora indicated that fish fed a diet with 300 mg kg β-glucan had higher bacterial richness and diversity as evaluated by Sobs, Chao, Ace, and Simpson indices, but no significant differences were found in the comparison groups. Furthermore, 300 mg kg β-glucan significantly increased the relative abundance of and decreased (mainly and ) and in largemouth bass intestinal microflora. The findings of this study provided new insights that will be valuable in future studies to elucidate the mechanism of immunity enhancement by β-glucan.
Topics: Animals; Transcriptome; Bass; Diet; Gene Expression Profiling
PubMed: 36591266
DOI: 10.3389/fimmu.2022.1086103 -
Fish & Shellfish Immunology Aug 2022As an inflammatory cytokine of the interleukin-20 (IL-20) subfamily, IL-20 has various functions in immune defenses, inflammatory diseases, tissue regeneration, cancer,...
Molecular identification and functional exploration of interleukin-20 in snakehead (Channa argus) involved in bacterial invasion and the proliferation of head kidney leukocytes.
As an inflammatory cytokine of the interleukin-20 (IL-20) subfamily, IL-20 has various functions in immune defenses, inflammatory diseases, tissue regeneration, cancer, and metabolism. Although the characteristics and functions of mammalian IL-20 have been clarified, those of fish IL-20 remain unclear. In this study, the IL-20 gene from the snakehead Channa argus (shIL-20) was cloned and functionally characterized. Similar to the IL-20 homologues of other species, the shIL-20 has a five exon/four intron structure in the coding region. The open reading frame of shIL-20 consists of 528 base pairs and encodes 175 amino acids (aa), including a signal peptide (aa 1-24) and a mature peptide (aa 25-175). The mature shIL-20 protein has six conserved cysteine residues, which occur in the IL-20 proteins of all species analyzed, and an additional cysteine residue (Cys-82) found only in the IL-20 proteins of several teleosts. The modeled tertiary structure of shIL-20 is similar with that of Homo sapiens IL-20. The shIL-20 was expressed constitutively in all the tissues analyzed, and its transcription was induced in the spleen and head kidney by Aeromonas schubertii and Nocardia seriolae in vivo and in head kidney leukocytes (HKLs) by lipoteichoic acid, lipopolysaccharide, and polyinosinic-polycytidylic acid in vitro. The recombinant shIL-20 protein induced the transcription of tumor necrosis factor α1 (TNF-α1), TNF-α2, IL-1β, and endogenous shIL-20, and promoted the proliferation of HKLs. In conclusion, these findings demonstrate that shIL-20 participates in the immune response to bacterial invasion and promotes leukocyte proliferation, offering new insights into the functions of fish IL-20 during pathogen invasion.
Topics: Animals; Bacteria; Cell Proliferation; Cysteine; Fish Diseases; Fish Proteins; Fishes; Head Kidney; Interleukins; Leukocytes; Mammals; Phylogeny
PubMed: 35810964
DOI: 10.1016/j.fsi.2022.07.005 -
Journal of Fish Diseases Aug 2022Motile Aeromonas septicemia (MAS), a disease caused by Aeromonas spp., is recognized as a major disease in freshwater aquaculture. This study aimed to investigate the...
Motile Aeromonas septicemia (MAS), a disease caused by Aeromonas spp., is recognized as a major disease in freshwater aquaculture. This study aimed to investigate the distribution and diversity of Aeromonas spp. and their antimicrobial susceptibility patterns. A total of 86 isolates of Aeromonas spp. were recovered from diseased freshwater fishes from 13 farms in Thailand. All isolates were identified using biochemical characteristics, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), polymerase chain reaction assays, and the gyrB gene sequence analysis. The result of MALDI-TOF MS showed 100% (86 isolates) accuracy at genus-level identification, and 88.4% (76 isolates) accuracy at species-level identification. Six species of Aeromonas were confirmed through nucleotide sequencing and phylogenetic analysis of the gyrB gene Aeromonas veronii (72.1%), Aeromonas jandaei (11.6%), Aeromonas schubertii (9.3%), Aeromonas diversa (3.5%), Aeromonas hydrophila (2.3%), and Aeromonas punctata (1.2%). Antimicrobial susceptibility tests for all isolates revealed resistance against amoxicillin (99%), ampicillin (98%), oxolinic acid (81.4%), oxytetracycline (77%), trimethoprim-sulfamethoxazole (24%), and enrofloxacin (21%). The multiple antibiotic resistance (MAR) index varied between 0.14 and 0.86, with MAR values more than 0.2 in 99% of isolates. Furthermore, four diverse multidrug-resistant (MDR) patterns were found among Aeromonas isolates. Our finding show that A. veronii is the most abundant species in Thai cultured freshwater fish with the highest MDR patterns.
Topics: Aeromonas; Animals; Anti-Bacterial Agents; Fish Diseases; Fishes; Fresh Water; Microbial Sensitivity Tests; Phylogeny; Thailand
PubMed: 35598068
DOI: 10.1111/jfd.13650 -
Veterinary World Jan 2022Antibiotic resistance has been a progressively documented problem, resulting in treatment failure in humans and animals. This study aimed to investigate the...
BACKGROUND AND AIM
Antibiotic resistance has been a progressively documented problem, resulting in treatment failure in humans and animals. This study aimed to investigate the antimicrobial susceptibility and virulence of extensively drug-resistant (XDR) spp. in wild and its surrounding seawater along the coastal road of Port Said, Egypt.
MATERIALS AND METHODS
Specimens were examined bacteriologically, confirmed biochemically, and tested for their sensitivity against 11 antimicrobial agents. Molecular confirmation of the obtained isolates by was performed, followed by the detection of antimicrobial resistance and virulence genes.
RESULTS
spp. was recovered from fish (44%) and water samples (36%). was the most prevalent identified strain, followed by , , and . Moreover, 90% of the tested isolates were multidrug-resistant (MDR), while 26.67% were XDR. Tested isolates were resistant to b-lactams and sulfonamides (100%), oxytetracycline (90%), and streptomycin (62.22%) but completely susceptible to cefotaxime. XDR isolates successfully amplified resistance genes (, and ()) but not the () gene, although there was phenotypic resistance to streptomycin on plates. All XDR isolates carry the cytotoxic enterotoxin gene (), but gene was detected in only one isolate (12.5%).
CONCLUSION
Data in this study provide a recent update and highlight the role of wild mullet and seawater as reservoirs for MDR and XDR spp. that may pose a risk to humans as food-borne infection or following direct contact.
PubMed: 35369605
DOI: 10.14202/vetworld.2022.55-64 -
Fish & Shellfish Immunology Apr 2022Interleukin-21 (IL-21), a crucial immune regulatory molecule, belongs to the common γ-chain family of type I cytokines, and exerts pleiotropic effects on multiple...
Characterization of snakehead (Channa argus) interleukin-21: Involvement in immune defense against two pathogenic bacteria, in leukocyte proliferation, and in activation of JAK-STAT signaling pathway.
Interleukin-21 (IL-21), a crucial immune regulatory molecule, belongs to the common γ-chain family of type I cytokines, and exerts pleiotropic effects on multiple immune cell types in mammals. However, the characteristics and functions of fish IL-21 remain unclear. To further investigate the molecular mechanism of IL-21 in teleosts, we first cloned and identified the IL-21 gene (designated shIL-21) of the snakehead (Channa argus). The full-length open reading frame of shIL-21 is 438 bp in length, and encodes a predicted protein of 145 amino acid residues. A sequence analysis showed that shIL-21 has the typical structural characteristics of other IL-21 proteins, containing four α-helices and four conserved cysteine residues. In a phylogenetic analysis, shIL-21 clustered within a subgroup of IL-21 proteins from other teleost species and shared its closest evolutionary relationship with that of Lates calcarifer. The expression analysis showed that shIL-21 was ubiquitously expressed in all the healthy snakehead tissues tested, albeit at different levels. After infection with Nocardia seriolae or Aeromonas schubertii, the relative expression of shIL-21 was mainly upregulated in the head kidney and spleen in vivo. Similarly, after stimulation with the three pathogen analogues lipoteichoic acid, lipopolysaccharides, and polyinosinic-polycytidylic acid, the expression of shIL-21 was also induced in head kidney leukocytes in vitro. A recombinant shIL-21 protein was expressed and purified, and promoted the proliferation of head kidney leukocytes, induced the expression of genes encoding critical signaling molecules in the Janus kinase (JAK) and signal transducer and activator of transcription (STAT) pathway, including JAK1, JAK3, STAT1, and STAT3, and induced the expression of endogenous shIL-21 and genes encoding several key proinflammatory cytokines (tumor necrosis factor-α, interferon-γ, and IL-1β). Taken together, these preliminary findings suggest that shIL-21 is involved in the immune defense against bacterial infection, in leukocyte proliferation, and in the activation of the JAK-STAT pathway. They thus extend the functional studies of IL-21 in teleosts.
Topics: Animals; Cell Proliferation; Fish Diseases; Fishes; Interleukins; Janus Kinases; Leukocytes; Mammals; Phylogeny; STAT Transcription Factors; Signal Transduction
PubMed: 35278639
DOI: 10.1016/j.fsi.2022.03.006 -
Microbiology Resource Announcements Jan 2022Aeromonas schubertii is a Gram-negative, rod-shaped bacterium. It is a rare species that has been reported in humans and aquatic animals. Here, we report the genome...
Aeromonas schubertii is a Gram-negative, rod-shaped bacterium. It is a rare species that has been reported in humans and aquatic animals. Here, we report the genome sequences of A. schubertii strains isolated from two mass mortality events in central Thailand that were associated with aquaculture of Asian seabass.
PubMed: 34989613
DOI: 10.1128/mra.01007-21 -
Pathogens (Basel, Switzerland) Aug 2021is the etiological pathogen of internal organ nodules in snakehead fish. Infections with produce a significant economic loss in aquaculture. Therefore, it is important...
is the etiological pathogen of internal organ nodules in snakehead fish. Infections with produce a significant economic loss in aquaculture. Therefore, it is important to examine the immune mechanisms by which snakeheads defend against infection. In this study, we established a hybrid snakehead infection model by intraperitoneal injection of that produced internal organ nodules. The splenic immune response of infected fish was examined at the transcriptome level by Illumina-seq analysis. Results showed 14,796 differentially expressed genes (DEGs) following infection, including 4441 up-regulated unigenes and 10,355 down-regulated unigenes. KEGG analysis showed 2084 DEGs to be involved in 192 pathways, 14 of which were immune-related. Twelve DEGs were used to validate quantitative real-time PCR results with RNA-seq data. Time-course expression analysis of six genes demonstrated modulation of the snakehead immune response by Furthermore, transcriptome analysis identified a substantial number of DEGs that were involved in the apoptosis signaling pathway. TUNEL analysis of infected spleens confirmed the presence of apoptotic cells. This study provided new information for a further understanding of the pathogenesis of in snakeheads, which can be used to prevent and possibly treat infections.
PubMed: 34451461
DOI: 10.3390/pathogens10080997 -
Frontiers in Public Health 2021Aeromonads are aquatic bacteria associated with frequent outbreaks of diarrhea in coastal Bangladesh, but their potential risks from environmental sources have remained...
Aeromonads are aquatic bacteria associated with frequent outbreaks of diarrhea in coastal Bangladesh, but their potential risks from environmental sources have remained largely unexplored. This study, over 2 years, examined homestead pond waters in the region for monthly dynamics and diversity of spp. The bacterial counts showed bi-modal annual growth peak, pre- and post-monsoon, strongly correlating ( < 0.0005) with temperature. Of 200 isolates characterized, bv. sobria (27%) was predominant among co-existent (20%), (17%), (13%), and three more. PCR screening of virulence-related genes identified 15 genotypes (I to XV), however, enterotoxigenicity in animal model was observed for five genotypes, ca. 18% (nine of 50) strains, prevalent in bv. sobria, , and . Pathogenic strains were distinguishable by possessing at least three of the major virulence genes: , and , together with accessory virulence factors. PFGE of I-digested genomic DNA revealed high genetic diversity and distant lineage of potentially toxigenic clones. Therefore, along with increased global warming, spp. having multi-factorial virulence potential in coastal ponds that serve as drinking water sources pose a potential health risk, and underscores the need for routine monitoring.
Topics: Aeromonas; Animals; Bangladesh; Ponds; Virulence; Water
PubMed: 34307285
DOI: 10.3389/fpubh.2021.692166