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13 pages, 10309 KiB

Open AccessArticle

Pathogenicity of Aeromonas veronii Isolated from Diseased Macrobrachium rosenbergii and Host Immune-Related Gene Expression Profiles

by Xiaojian Gao, Zhen Chen, Zirui Zhang, Qieqi Qian, Anting Chen, Lijie Qin, Xinzhe Tang, Qun Jiang and Xiaojun Zhang

Microorganisms 2024, 12(4), 694; https://doi.org/10.3390/microorganisms12040694 - 29 Mar 2024

Cited by 1 | Viewed by 1265

Abstract

Aeromonas veronii is widespread in aquatic environments and is responsible for infecting various aquatic animals. In this study, a dominant strain was isolated from the hepatopancreas of diseased Macrobrachium rosenbergii and was named JDM1-1. According to its morphological, physiological, and biochemical characteristics and [...] Read more.

Aeromonas veronii is widespread in aquatic environments and is responsible for infecting various aquatic animals. In this study, a dominant strain was isolated from the hepatopancreas of diseased Macrobrachium rosenbergii and was named JDM1-1. According to its morphological, physiological, and biochemical characteristics and molecular identification, isolate JDM1-1 was identified as A. veronii. The results of artificial challenge showed isolate JDM1-1 had high pathogenicity to M. rosenbergii with an LD₅₀ value of 8.35 × 10⁵ CFU/mL during the challenge test. Histopathological analysis revealed severe damage in the hepatopancreas and gills of the diseased prawns, characterized by the enlargement of the hepatic tubule lumen and gaps between the tubules as well as clubbing and degeneration observed at the distal end of the gill filament. Eight virulence-related genes, namely aer, ompA, lip, tapA, hlyA, flgA, flgM, and flgN, were screened by PCR assay. In addition, virulence factor detection showed that the JDM1-1 isolate produced lipase, lecithinase, gelatinase, and hemolysin. Furthermore, the mRNA expression profiles of immune-related genes of M. rosenbergii following A. veronii infection, including ALF1, ALF2, Crustin, C-lectin, and Lysozyme, were assessed, and the results revealed a significant upregulation in the hepatopancreas and intestines at different hours post infection. This study demonstrates that A. veronii is a causative agent associated with massive die-offs of M. rosenbergii and contributes valuable insights into the pathogenesis and host defense mechanisms of A. veronii invasion. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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20 pages, 32847 KiB

Open AccessArticle

Coinfection of Cage-Cultured Spotted Sea Bass (Lateolabrax maculatus) with Vibrio harveyi and Photobacterium damselae subsp. piscicida Associated with Skin Ulcer

by Dandan Zhou, Binzhe Zhang, Yuchen Dong, Xuepeng Li and Jian Zhang

Microorganisms 2024, 12(3), 503; https://doi.org/10.3390/microorganisms12030503 - 29 Feb 2024

Cited by 1 | Viewed by 1509

Abstract

Spotted sea bass (Lateolabrax maculatus) is a high-economic-value aquacultural fish widely distributed in the coastal and estuarine areas of East Asia. In August 2020, a sudden outbreak of disease accompanied by significant mortality was documented in L. maculatus reared in marine [...] Read more.

Spotted sea bass (Lateolabrax maculatus) is a high-economic-value aquacultural fish widely distributed in the coastal and estuarine areas of East Asia. In August 2020, a sudden outbreak of disease accompanied by significant mortality was documented in L. maculatus reared in marine cage cultures located in Nanhuang island, Yantai, China. Two coinfected bacterial strains, namely, NH-LM1 and NH-LM2, were isolated from the diseased L. maculatus for the first time. Through phylogenetic tree analysis, biochemical characterization, and genomic investigation, the isolated bacterial strains were identified as Vibrio harveyi and Photobacterium damselae subsp. piscicida, respectively. The genomic analysis revealed that V. harveyi possesses two circular chromosomes and six plasmids, while P. damselae subsp. piscicida possesses two circular chromosomes and two plasmids. Furthermore, pathogenic genes analysis identified 587 and 484 genes in V. harveyi and P. damselae subsp. piscicida, respectively. Additionally, drug-sensitivity testing demonstrated both V. harveyi and P. damselae subsp. piscicida exhibited sensitivity to chloramphenicol, ciprofloxacin, ofloxacin, orfloxacin, minocycline, doxycycline, tetracycline, and ceftriaxone. Moreover, antibiotic resistance genes were detected in the plasmids of both strains. Extracellular product (ECP) analysis demonstrated that both V. harveyi and P. damselae subsp. piscicida can produce hemolysin and amylase, while V. harveyi additionally can produce caseinase and esterase. Furthermore, infected fish displayed severe histopathological alterations, including infiltration of lymphocytes, cellular degeneration and necrosis, and loose aggregation of cells. Artificial infection assays determined that the LD₅₀ of P. damselae subsp. piscicida was 3 × 10⁵ CFU/g, while the LD₅₀ of V. harveyi was too low to be accurately evaluated. Furthermore, the dual infection of V. harveyi and P. damselae subsp. piscicida elicits a more rapid and pronounced mortality rate compared to single challenge, thereby potentially exacerbating the severity of the disease through synergistic effects. Ultimately, our findings offer compelling evidence for the occurrence of coinfections involving V. harveyi and P. damselae subsp. piscicida in L. maculatus, thereby contributing to the advancement of diagnostic and preventative measures for the associated disease. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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17 pages, 4543 KiB

Open AccessArticle

The Mutation of the DNA-Binding Domain of Fur Protein Enhances the Pathogenicity of Edwardsiella piscicida via Inducing Overpowering Pyroptosis

by Mimi Niu, Zhihai Sui, Guoquan Jiang, Ling Wang, Xuemei Yao and Yonghua Hu

Microorganisms 2024, 12(1), 11; https://doi.org/10.3390/microorganisms12010011 - 20 Dec 2023

Cited by 2 | Viewed by 1169

Abstract

Edwardsiella piscicida is an important fish pathogen with a broad host that causes substantial economic losses in the aquaculture industry. Ferric uptake regulator (Fur) is a global transcriptional regulator and contains two typical domains, the DNA-binding domain and dimerization domain. In a previous [...] Read more.

Edwardsiella piscicida is an important fish pathogen with a broad host that causes substantial economic losses in the aquaculture industry. Ferric uptake regulator (Fur) is a global transcriptional regulator and contains two typical domains, the DNA-binding domain and dimerization domain. In a previous study, we obtained a mutant strain of full-length fur of E. piscicida, TX01Δfur, which displayed increased siderophore production and stress resistance factors and decreased pathogenicity. To further reveal the regulatory mechanism of Fur, the DNA-binding domain (N-terminal) of Fur was knocked out in this study and the mutant was named TX01Δfur2. We found that TX01Δfur2 displayed increased siderophore production and enhanced adversity tolerance, including a low pH, manganese, and high temperature stress, which was consistent with the phenotype of TX01Δfur. Contrary to TX01Δfur, whose virulence was weakened, TX01Δfur2 displayed an ascended invasion of nonphagocytic cells and enhanced destruction of phagocytes via inducing overpowering or uncontrollable pyroptosis, which was confirmed by the fact that TX01Δfur2 induced higher levels of cytotoxicity, IL-1β, and p10 in macrophages than TX01. More importantly, TX01Δfur2 displayed an increased global virulence to the host, which was confirmed by the result that TX01Δfur2 caused higher lethality outcomes for healthy tilapias than TX01. These results demonstrate that the mutation of the Fur N-terminal domain augments the resistance level against the stress and pathogenicity of E. piscicida, which is not dependent on the bacterial number in host cells or host tissues, although the capabilities of biofilm formation and the motility of TX01Δfur2 decline. These interesting findings provide a new insight into the functional analysis of Fur concerning the regulation of virulence in E. piscicida and prompt us to explore the subtle regulation mechanism of Fur in the future. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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11 pages, 4552 KiB

Open AccessArticle

Vibrio splendidus AJ01 Promotes Pathogenicity via L-Glutamic Acid

by Ya Li, Weibo Shi and Weiwei Zhang

Microorganisms 2023, 11(9), 2333; https://doi.org/10.3390/microorganisms11092333 - 17 Sep 2023

Cited by 2 | Viewed by 1326

Abstract

Vibrio splendidus is a pathogen that infects a wide range of hosts, especially the sea cucumber species Apostichopus japonicus. Previous studies showed that the level of L-glutamic acid (L-Glu) significantly increased under heat stress, and it was found to be one of [...] Read more.

Vibrio splendidus is a pathogen that infects a wide range of hosts, especially the sea cucumber species Apostichopus japonicus. Previous studies showed that the level of L-glutamic acid (L-Glu) significantly increased under heat stress, and it was found to be one of the best carbon sources used by V. splendidus AJ01. In this study, the effects of exogenous L-Glu on the coelomocyte viability, tissue status, and individual mortality of sea cucumbers were analyzed. The results showed that 10 mM of L-Glu decreased coelomocyte viability and increased individual mortality, with tissue rupture and pyknosis, while 0.1 mM of L-Glu slightly affected the survival of sea cucumbers without obvious damage at the cellular and tissue levels. Transcriptomic analysis showed that exogenous L-Glu upregulated 343 and downregulated 206 genes. Gene Ontology (GO) analysis showed that differentially expressed genes (DEGs) were mainly enriched in signaling and membrane formation, while a Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that DEGs were significantly enriched in the upregulated endocytosis and downregulated lysosomal pathways. The coelomocyte viability further decreased by 20% in the simultaneous presence of exogenous L-Glu and V. splendidus AJ01 compared with that in the presence of V. splendidus AJ01 infection alone. Consequently, a higher sea cucumber mortality was also observed in the presence of exogenous L-Glu challenged by V. splendidus AJ01. Real-time reverse transcriptase PCR showed that L-Glu specifically upregulated the expression of the fliC gene coding the subunit protein of the flagellar filament, promoting the swimming motility activity of V. splendidus. Our results indicate that L-Glu should be kept in a state of equilibrium, and excess L-Glu at the host–pathogen interface prompts the virulence of V. splendidus via the increase of bacterial motility. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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16 pages, 4916 KiB

Open AccessArticle

The Thioredoxin System in Edwardsiella piscicida Contributes to Oxidative Stress Tolerance, Motility, and Virulence

by Jiaojiao He, Su Liu, Qingjian Fang, Hanjie Gu and Yonghua Hu

Microorganisms 2023, 11(4), 827; https://doi.org/10.3390/microorganisms11040827 - 24 Mar 2023

Cited by 4 | Viewed by 1820

Abstract

Edwardsiella piscicida is an important fish pathogen that causes substantial economic losses. In order to understand its pathogenic mechanism, additional new virulence factors need to be identified. The bacterial thioredoxin system is a major disulfide reductase system, but its function is largely unknown [...] Read more.

Edwardsiella piscicida is an important fish pathogen that causes substantial economic losses. In order to understand its pathogenic mechanism, additional new virulence factors need to be identified. The bacterial thioredoxin system is a major disulfide reductase system, but its function is largely unknown in E. piscicida. In this study, we investigated the roles of the thioredoxin system in E. piscicida (named TrxB_Ep, TrxA_Ep, and TrxC_Ep, respectively) by constructing a correspondingly markerless in-frame mutant strain: ΔtrxB, ΔtrxA, and ΔtrxC, respectively. We found that (i) TrxB_Ep is confirmed as an intracellular protein, which is different from the prediction made by the Protter illustration; (ii) compared to the wild-type strain, ΔtrxB exhibits resistance against H₂O₂ stress but high sensitivity to thiol-specific diamide stress, while ΔtrxA and ΔtrxC are moderately sensitive to both H₂O₂ and diamide conditions; (iii) the deletions of trxB_Ep, trxA_Ep, and trxC_Ep damage E. piscicida’s flagella formation and motility, and trxB_Ep plays a decisive role; (iv) deletions of trxB_Ep, trxA_Ep, and trxC_Ep substantially abate bacterial resistance against host serum, especially trxB_Ep deletion; (v) trxA_Ep and trxC_Ep, but not trxB_Ep, are involved in bacterial survival and replication in phagocytes; (vi) the thioredoxin system participates in bacterial dissemination in host immune tissues. These findings indicate that the thioredoxin system of E. piscicida plays an important role in stress resistance and virulence, which provides insight into the pathogenic mechanism of E. piscicida. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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16 pages, 2801 KiB

Open AccessArticle

Effect of the Type VI Secretion System Secreted Protein Hcp on the Virulence of Aeromonas salmonicida

by Hongyan Cai, Jiaying Yu, Ying Qiao, Ying Ma, Jiang Zheng, Mao Lin, Qingpi Yan and Lixing Huang

Microorganisms 2022, 10(12), 2307; https://doi.org/10.3390/microorganisms10122307 - 22 Nov 2022

Cited by 9 | Viewed by 2187

Abstract

Aeromonas salmonicida, a psychrophilic bacterial pathogen, is widely distributed in marine freshwater, causing serious economic losses to major salmon farming areas in the world. At present, it is still one of the most important pathogens threatening salmon farming. Hcp (haemolysin-coregulated protein) is [...] Read more.

Aeromonas salmonicida, a psychrophilic bacterial pathogen, is widely distributed in marine freshwater, causing serious economic losses to major salmon farming areas in the world. At present, it is still one of the most important pathogens threatening salmon farming. Hcp (haemolysin-coregulated protein) is an effector protein in the type-VI secretion system (T6SS), which is secreted by T6SS and functions as its structural component. The results of our previous genomic sequencing showed that hcp existed in the mesophilic A. salmonicida SRW-OG1 isolated from naturally infected Epinephelus coioides. To further explore the role of Hcp in A. salmonicida SRW-OG1, we constructed an hcp-RNAi strain and verified its effect on the virulence of A. salmonicida. The results showed that compared with the wild strain, the hcp-RNAi strain suffered from different degrees of decreased adhesion, growth, biofilm formation, extracellular product secretion, and virulence. It was suggested that hcp may be an important virulence gene of A. salmonicida SRW-OG1. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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12 pages, 3822 KiB

Open AccessArticle

Pathogenicity of Aeromonas veronii Causing Mass Mortality of Largemouth Bass (Micropterus salmoides) and Its Induced Host Immune Response

by Xinhai Zhu, Qieqi Qian, Congcong Wu, Yujie Zhu, Xiaojian Gao, Qun Jiang, Jun Wang, Guoxing Liu and Xiaojun Zhang

Microorganisms 2022, 10(11), 2198; https://doi.org/10.3390/microorganisms10112198 - 6 Nov 2022

Cited by 18 | Viewed by 2729

Abstract

Aeromonas veronii is as an important opportunist pathogen of many aquatic animals, which is wildly distributed in various aquatic environments. In this study, a dominant bacterium GJL1 isolated from diseased M. salmoides was identified as A. veronii according to the morphological, physiological, and [...] Read more.

Aeromonas veronii is as an important opportunist pathogen of many aquatic animals, which is wildly distributed in various aquatic environments. In this study, a dominant bacterium GJL1 isolated from diseased M. salmoides was identified as A. veronii according to the morphological, physiological, and biochemical characteristics, as well as molecular identification. Detection of the virulence genes showed the isolate GJL1 carried outer membrane protein A (ompA), flagellin (flgA, flgM, flgN), aerolysin (aer), cytolytic enterotoxin (act), DNases (exu), and hemolysin (hly), and the isolate GJL1 also produced caseinase, lipase, gelatinase, and hemolysin. The virulence of strain GJL1 was confirmed by experimental infection; the median lethal dosage (LD₅₀) of the GJL1 for largemouth bass was 3.6 × 10⁵ CFU/mL, and histopathological analysis revealed that the isolate could cause obvious inflammatory responses in M. salmoides. Additionally, the immune-related gene expression in M. salmoides was evaluated, and the results showed that IgM, HIF-1α, Hep-1, IL-15, TGF-β1, and Cas-3 were significantly upregulated after A. veronii infection. Our results indicated that A. veronii was an etiological agent causing the mass mortality of M. salmoides, which contributes to understanding the immune response of M. salmoides against A. veronii infection. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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16 pages, 3232 KiB

Open AccessArticle

The Early Peritoneal Cavity Immune Response to Vibrio Anguillarum Infection and to Inactivated Bacterium in Olive Flounder (Paralichthys olivaceus)

by Xueyan Shi, Heng Chi, Yuanyuan Sun, Xiaoqian Tang, Jing Xing, Xiuzhen Sheng and Wenbin Zhan

Microorganisms 2022, 10(11), 2175; https://doi.org/10.3390/microorganisms10112175 - 2 Nov 2022

Cited by 7 | Viewed by 2021

Abstract

The peritoneal cavity plays an important role in the immune response, and intraperitoneal administration is an ideal vaccination route in fish. However, immune responses in the peritoneal cavity of teleost fish are still not completely characterized. This study characterized the morphology of peritoneal [...] Read more.

The peritoneal cavity plays an important role in the immune response, and intraperitoneal administration is an ideal vaccination route in fish. However, immune responses in the peritoneal cavity of teleost fish are still not completely characterized. This study characterized the morphology of peritoneal cavity cells (PerC cells) and their composition in flounder (Paralichthys olivaceus). Flow cytometric analysis of the resident PerC cells revealed two populations varying in granularity and size. One population, approximately 15.43% ± 1.8%, was smaller with a lower granularity, designated as lymphocytes. The other population of the cells, about 78.17% ± 3.52%, was larger with higher granularity and was designated as myeloid cells. The results of cytochemical staining and transmission electron microscopy indicated that peritoneal cavity in flounder normally contains a resident population of leukocytes dominated by granulocytes, macrophages, dendritic cells, and lymphocytes. The percentages of IgM⁺, CD4⁺, G-CSFR⁺, MHCII⁺, and CD83⁺ leukocytes among PerC cells determined by flow cytometry were 3.13% ± 0.4%, 2.83% ± 0.53%, 21.12% ± 1.44%, 27.11% ± 3.30%, and 19.64% ± 0.31%, respectively. Further, the changes in IgM⁺, CD4⁺, G-CSFR⁺, MHCII⁺, and CD83⁺ leukocytes in flounder after Vibrio anguillarum infection and immunization were compared. The composition changed rapidly after the infection or vaccination treatment and included two stages, a non-specific stage dominated by phagocytes and a specific immune stage dominated by lymphocytes. Due to the virulence effectors of bacteria, the infected group exhibited a more intense and complicated PerC cells immune response than that of the immunization group. Following our previous study, this is the first report on the morphology and composition of PerC cells and the early activation of PerC cells in flounder response to V. anguillarum infection and vaccination. Full article

(This article belongs to the Special Issue Immunotoxic Factors Promoting Infectious Diseases in Cultured Marine Animals)

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