Antimicrobial activity of coral-associated beneficial bacteria against coral disease-causing microbial pathogens.

Microbial infection of immune-compromised corals influences disease severity, resulting in coral mortality. However, coral-associated beneficial bacteria are known to produce antimicrobial compounds that prevent the growth of potential pathogens and invading microbes. Hence, beneficial bacteria associated with coral Porites lutea were isolated and antimicrobial protein and bioactive secondary metabolites were extracted and tested for their antimicrobial activity against putative prokaryotic and eukaryotic coral pathogens. Bioactive secondary metabolites exhibited remarkable antagonism against various coral pathogens such as Serratia marcescens, Vibrio species, and Aspergillus sydowii. Besides, the metabolites of Cobetia marina, Cobetia amphilecti, Pseudoalteromonas neustonica, and Virgibacillus halodenitrificans manifested notable inhibition against the protozoan ciliates (Uronema marinum, Holosticha diademata, Cohnilembus verminus, and Euplotes vannus) and zooplankton that are known to be involved in the secondary pathogenesis in coral diseased lesion progression. Thus, the present study may benefit in understanding coral-associated beneficial bacteria for their antagonistic interactions with microbial pathogens, as well as their potential involvement in reducing coral disease severity.

Characterization of the defense properties of healthy and diseased coral mucus.

The surface mucus layer of corals aids in feeding, silt removal, and defense against pathogens. However, first line of defense of secreted mucus of corals against tissue damage generated through pathogen or environmental factors is poorly understood. Hence, we used various methods such as a well diffusion assay and tests for quorum quenching, free radical scavenging, antioxidant enzyme expression and phenoloxidase (PO) activity to determine the mucus defense properties using mucus of healthy and diseased Porites sp. and Acropora sp. Interestingly the coral mucus showed antimicrobial activity against coral pathogens such as bacteria and protozoan ciliates. Inhibition of the N-Acyl homoserine lactone (AHL) molecule suggests quorum quenching. Free radical scavenging of mucus was screened using hydrogen peroxide, hydroxyl radical, and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) methods, which was found significantly more in diseased corals (p < 0.05). Antioxidant enzymes superoxide dismutase (SOD), catalase, and peroxidase activity were observed in both the diseased and healthy coral mucus. The presence of serine and metalloproteases was also detected in coral mucus. Further, phenoloxidase (PO) activity was highest in diseased coral mucus affected by pink line syndrome and white patch Acropora sp. disease than the healthy coral mucus. Thus, the present study of antimicrobial properties, antioxidant enzymes, and quorum quenching properties in coral mucus may aid in understanding the corals defense and survival against pathogens and any abiotic stress.

Possible beneficial interactions of ciliated protozoans with coral health and resilience.

Microbial interactions contribute significantly to coral health in the marine environment. Most beneficial associations have been described with their bacterial communities, but knowledge of beneficial associations between protozoan ciliates and corals is still lacking. Ciliates are important bacterial predators and provide nutrition to higher trophic-level organisms. The mucus secreted by corals and the microenvironment of the coral surface layer attract ciliates based on their food preferences. The mixotrophic and heterotrophic ciliates play a major role in nutrient cycling by increasing nitrogen, phosphorus, and extractable sulfur, which can enhance the proliferation of coral beneficial microbe. Besides, bacterial predator ciliates reduce the pathogenic bacterial population that infects the coral and also act as bioindicators for assessing the toxicity of the reef ecosystem. Thus, these ciliates can be used as a beneficial partner in influencing coral health and resilience under various stress conditions. Herein, we explore the urgent need to understand the complex beneficial interactions of ciliates that may occur in the coral reef ecosystem.

Manipulating Galectin Expression in Zebrafish (Danio rerio).

Techniques for disrupting gene expression are invaluable tools for the analysis of the biological role of a gene product. Because of its genetic tractability and multiple advantages over conventional mammalian models, the zebrafish (Danio rerio) is recognized as a powerful system for gaining new insight into diverse aspects of human health and disease. Among the multiple mammalian gene families for which the zebrafish has shown promise as an invaluable model for functional studies, the galectins have attracted great interest due to their participation in early development, regulation of immune homeostasis, and recognition of microbial pathogens. Galectins are ß-galactosyl-binding lectins with a characteristic sequence motif in their carbohydrate recognition domains (CRDs), that constitute an evolutionary conserved family ubiquitous in eukaryotic taxa. Galectins are emerging as key players in the modulation of many important pathological processes, which include acute and chronic inflammatory diseases, autoimmunity and cancer, thus making them potential molecular targets for innovative drug discovery. Here, we provide a review of the current methods available for the manipulation of gene expression in the zebrafish, with a focus on gene knockdown [morpholino (MO)-derived antisense oligonucleotides] and knockout (CRISPR-Cas) technologies.

Ciliated protozoan occurrence and association in the pathogenesis of coral disease.

Various microbial infections have significantly contributed to disease progression leading to the mortality of corals. However, the holobiont and the external surfaces of coral, including the secreted mucus, provide a varied microenvironment that attracts ciliates based on their feeding preferences. Besides, some ciliates (e.g., Philasterine scuticociliate) may enter through the injuries or lesions on corals or through their indirect interactions with other types of microbes that influence coral health. Thus, ciliates occurrence and association are described with 12 different diseases worldwide. White syndrome disease lesions have diverse ciliate associations, and higher ciliate diversity was observed with diseased genera Acropora. Also, it was described, about sixteen ciliate species ingest coral Symbiodiniaceae and histophagous ciliates for coral tissue loss as secondary invaders. However, the ciliates nature of association with the coral disease remains unclear for primary or opportunistic secondary pathogenicity. Herein, we explore the urgent need to understand the complex interactions of ciliates in coral health.

Characterization of coral associated ciliates and their interactions with disease lesion progression of Indian Scleractinian corals.

Ciliates associated with advanced diseased lesions of Acropora sp. and Porites sp. in the field were isolated and characterised using microscopic and molecular analysis. The identified dominant coral-associated ciliates as Holosticha sp. and Cohnilembusverminus was propagated in vitro and taken for further study. Ciliates high cell numbers with substrate containing bacteria-free mucus confirms the feeding preference for nutrients in mucus instead of bacteria. Therefore, fatty acid composition of the coral mucus was analysed and noted for the different composition levels of SAFA, MUFA and PUFA in both the genera. This suggests the possible feed specific interactions of ciliates with coral mucus and tissues. Conversely, Holosticha sp. was observed for invading the host cells for its voracious ingestion of Symbiodiniaceae cells and tissues. Moreover, the aquarium based investigation revealed that the ciliates migrate to the injured and early disease signs of corals enhancing the tissue loss and disease lesion progression. Thus, our results indicate that the ciliates interact with the immunocompromised disease corals and play a major role in progression of disease lesions leading to rapid coral mortality.

Diseases and pathogens of marine invertebrate corals in Indian reefs.

Diseases in marine invertebrate corals have been reported worldwide and have been associated with infection by various microbial pathogens that cause massive mortality. Several bacterial species, especially Vibrio species but also members of the cyanobacteria, fungi, viruses, and protists, are described as important pathogens associated with coral disease and mortality. The present work provides an updated overview of main diseases and implicated microbial species affecting corals in Indian reefs. Further study on pathogen diversity, classification, spread and environmental factors on pathogen-host interactions may contribute a better understanding of the coral diseases.

The zebrafish galectins Drgal1-L2 and Drgal3-L1 bind in vitro to the infectious hematopoietic necrosis virus (IHNV) glycoprotein and reduce viral adhesion to fish epithelial cells.

The infectious hematopoietic necrosis virus (IHNV; Rhabdoviridae, Novirhabdovirus) infects teleost fish, such as salmon and trout, and is responsible for significant losses in the aquaculture industry and in wild fish populations. Although IHNV enters the host through the skin at the base of the fins, the viral adhesion and entry mechanisms are not fully understood. In recent years, evidence has accumulated in support of the key roles played by protein-carbohydrate interactions between host lectins secreted to the extracellular space and virion envelope glycoproteins in modulating viral adhesion and infectivity. In this study, we assessed in vitro the potential role(s) of zebrafish (Danio rerio) proto type galectin-1 (Drgal1-L2) and a chimera galectin-3 (Drgal3-L1) in IHNV adhesion to epithelial cells. Our results suggest that the extracellular Drgal1-L2 and Drgal3-L1 interact directly and in a carbohydrate-dependent manner with the IHNV glycosylated envelope and glycans on the epithelial cell surface, significantly reducing viral adhesion.

Infection and pathogenecity of Myroides odoratimimus (NIOCR-12) isolated from the gut of grey mullet (Mugil cephalus (Linnaeus, 1758)).

Myroides sp. are gram negative aerobes and known for its oppurtunistic pathogenicity in humans. In the present study, Myroides odoratimimus isolated from the gut of Mugil cephalus showed potential infectivity to the experimental grey mullet and acted as an ultimate pathogen with significant symptoms. Furthermore, the inoculum isolated from the infected fishes were cultured and the selected colonies were reisolated and reinjected into healthy juveniles of M. cephalus. Characterizations of the re-isolated bacteria were the same as those of the isolated M. odoratimimus from naturally infected mullet. The median lethal dose (LD50) of the bacteria was 3 × 10(6) CFU fish(-1). In order to assay the accuracy of infection, the production of reactive oxygen species (ROS), the respiratory burst activity of blood leukocytes of mullet before and after challenge was measured, an indicator of the innate immune system. The mullet infection increased the respiratory burst activity and superoxide dismutase activity. In addition, the innate immune response of TLR 9 expression against M. odoratimimus infection and CpG ODN treatment in disease model, zebrafish confirms the M. odoratimimus infection and pathogenicity.

Manipulating galectin expression in zebrafish (Danio rerio).

Techniques for disrupting gene expression are invaluable tools for the analysis of the biological role(s) of a gene product. Because of its genetic tractability and multiple advantages over conventional mammalian models, the zebrafish (Danio rerio) is recognized as a powerful system for gaining new insight into diverse aspects of human health and disease. Among the multiple mammalian gene families for which the zebrafish has shown promise as an invaluable model for functional studies, the galectins have attracted great interest due to their participation in early development, regulation of immune homeostasis, and recognition of microbial pathogens. Galectins are ß-galactosyl-binding lectins with a characteristic sequence motif in their carbohydrate recognition domains (CRDs), which comprise an evolutionary conserved family ubiquitous in eukaryotic taxa. Galectins are emerging as key players in the modulation of many important pathological processes, which include acute and chronic inflammatory diseases, autoimmunity and cancer, thus making them potential molecular targets for innovative drug discovery. Here, we provide a review of the current methods available for the manipulation of gene expression in the zebrafish, with a focus on gene knockdown [morpholino (MO)-derived antisense oligonucleotides] and knockout (CRISPR-Cas) technologies.

A role for antioxidants in acclimation of marine derived pathogenic fungus (NIOCC 1) to salt stress.

Salinity tolerance a key factor helps in understanding the ionic homeostasis in general, which is a fundamental cellular phenomenon in all living cells. Here, a marine derived pathogenic fungus was examined for its adaptation under salt stress using antioxidant properties. The aqueous extracts of halophilic fungus exhibited different levels of antioxidant activity in all the in vitro tests such as α,α-diphenyl-ß-picrylhydrazyl (DPPH(·)), Hydroxyl Radical Scavenging Assay (HRSA), Metal chelating assay and ß-carotene-linoleic acid model system. The antioxidant capacity of marine fungus exposed to high salt condition showed an increase in activity. In addition, the production of intra and extracellular antioxidant enzymes of the fungus at various salt stresses were analyzed and discussed for their possible role in the stress mechanism. The marine derived fungus was identified as Phialosimplex genus, which is associated with infections in dogs. Thus the present study elucidates that the scavenging activity is one of the protective mechanisms developed in the fungus to avoid the deleterious effect of salt stress. In addition, the study also helps in understanding how the pathogenic fungus tackles the oxidative burst i.e. hypersensitivity reaction performed by host to kill the pathogens.