Study Of Host-pathogen Interactions Research Articles

Take my breath away: studying pathogen invasion of the human lung using primary tissue models.

The human pulmonary environment is complex, containing a matrix of cells, including fibroblasts, epithelial cells, interstitial macrophages, alveolar macrophages and neutrophils. When confronted with foreign material or invading pathogens, these cells mount a robust response. Nevertheless, many bacterial pathogens with an intracellular lifecycle stage exploit this environment for replication and survival. These include, but are not limited to, Coxiella burnetii, Legionella pneumophila, Yersinia pestis, Mycobacterium tuberculosis and Staphylococcus aureus. Currently, few human disease-relevant model systems exist for studying host-pathogen interactions during these bacterial infections in the lung. Here, we present two novel infection platforms, human alveolar macrophages (hAMs) and human precision-cut lung slices (hPCLS), along with an up-to-date synopsis of research using said models. Additionally, alternative uses for these systems in the absence of pathogen involvement are presented, such as tissue banking and further characterization of the human lung environment. Overall, hAMs and hPCLS allow novel human disease-relevant investigations that other models, such as cell lines and animal models, cannot completely provide.

Influence of Culture Media and Temperature on Growth and Sporulation of Ceratocystis lukuohia

Ceratocystis lukuohia is one of two newly described pathogens of Metrosideros polymorpha (‘ōhi‘a) causing Rapid ‘Ōhi‘a Death, a phenomenon that is devastating sections of native forest across the state of Hawai‘i, USA. Ceratocystis lukuohia research has increased as the threat of the pathogen has become more apparent, resulting in a need for efficient production of fungal cultures to meet research demands. Therefore, the temperature and medium-dependent mycelium growth and spore production of three isolates, one from the outbreak area of Leilani Estates and two from beyond the area, were assessed in culture. Fungal growth, measured as mycelium diameter, did not differ for the three isolates after 7 days of incubation. Optimal growth temperatures were 25–30 °C on 10% V8 agar and 25 °C on malt yeast extract agar (MYEA). Spore production, as assessed in one isolate, was most abundant at 25 °C and on MYEA, followed by 10% V8, and MEA. Mycelium growth and spore production on 10% V8 were less variable compared to the other media and, while mean spore production was highest on MYEA, the test isolate produced at least 1 × 106 spores/mL on 10% V8 agar, a commonly used spore concentration for inoculation experiments. Therefore, we recommend growing C. lukuohia at 25 °C on 10% V8 for inoculum production. These results contribute to the growing knowledge base of this newly discovered fungal pathogen and suggest a standardized methodology for propagating C. lukuohia for use in host-pathogen interaction studies.

Advances in modelling the human microbiome-gut-brain axis in vitro.

The human gut microbiome has emerged as a key player in the bidirectional communication of the gut–brain axis, affecting various aspects of homeostasis and pathophysiology. Until recently, the majority of studies that seek to explore the mechanisms underlying the microbiome–gut–brain axis cross-talk, relied almost exclusively on animal models, and particularly gnotobiotic mice. Despite the great progress made with these models, various limitations, including ethical considerations and interspecies differences that limit the translatability of data to human systems, pushed researchers to seek for alternatives. Over the past decades, the field of in vitro modelling of tissues has experienced tremendous growth, thanks to advances in 3D cell biology, materials, science and bioengineering, pushing further the borders of our ability to more faithfully emulate the in vivo situation. The discovery of stem cells has offered a new source of cells, while their use in generating gastrointestinal and brain organoids, among other tissues, has enabled the development of novel 3D tissues that better mimic the native tissue structure and function, compared with traditional assays. In parallel, organs-on-chips technology and bioengineered tissues have emerged as highly promising alternatives to animal models for a wide range of applications. Here, we discuss how recent advances and trends in this area can be applied in host–microbe and host–pathogen interaction studies. In addition, we highlight paradigm shifts in engineering more robust human microbiome-gut-brain axis models and their potential to expand our understanding of this complex system and hence explore novel, microbiome-based therapeutic approaches.

Gnotobiotic rainbow trout (Oncorhynchus mykiss) model reveals endogenous bacteria that protect against Flavobacterium columnare infection.

The health and environmental risks associated with antibiotic use in aquaculture have promoted bacterial probiotics as an alternative approach to control fish infections in vulnerable larval and juvenile stages. However, evidence-based identification of probiotics is often hindered by the complexity of bacteria-host interactions and host variability in microbiologically uncontrolled conditions. While these difficulties can be partially resolved using gnotobiotic models harboring no or reduced microbiota, most host-microbe interaction studies are carried out in animal models with little relevance for fish farming. Here we studied host-microbiota-pathogen interactions in a germ-free and gnotobiotic model of rainbow trout (Oncorhynchus mykiss), one of the most widely cultured salmonids. We demonstrated that germ-free larvae raised in sterile conditions displayed no significant difference in growth after 35 days compared to conventionally-raised larvae, but were extremely sensitive to infection by Flavobacterium columnare, a common freshwater fish pathogen causing major economic losses worldwide. Furthermore, re-conventionalization with 11 culturable species from the conventional trout microbiota conferred resistance to F. columnare infection. Using mono-re-conventionalized germ-free trout, we identified that this protection is determined by a commensal Flavobacterium strain displaying antibacterial activity against F. columnare. Finally, we demonstrated that use of gnotobiotic trout is a suitable approach for the identification of both endogenous and exogenous probiotic bacterial strains protecting teleostean hosts against F. columnare. This study therefore establishes an ecologically-relevant gnotobiotic model for the study of host-pathogen interactions and colonization resistance in farmed fish.

Functional proteoliposome-like structure derived from simultaneous evisceration and enucleation of T-lymphoblastoid A3R5.7 cells: A top-down story

Structurally-reduced cells and cell-derived structures are powerful tools for membrane studies. Using this approach, we probed whether a cell, without its nucleus and cytoplasm, is still capable of undergoing CD4-mediated membrane fusion. For this, we needed a cell-derived structure, akin to a giant liposome functionalised with CD4 and chemokine receptors. We present a method for the simultaneous removal of cytoplasmic and nuclear material from cells presenting CD4, CCR5, and CXCR4, using Colcemid treatment followed by hypotonic cytolysis, and then enriched using preparative flow cytometry. We show that the resultant cell membrane remains intact, retains presentation of CD4, CCR5, and CXCR4, and is still capable of CD4-mediated membrane fusion with a target cell. Finally, we detail how this protocol was developed, as well as how such samples should be handled for storage and assays. We envision the use of such systems for host-pathogen interaction studies, and the development of targeted delivery vehicles.

A Paradigm Gap in Host-Pathogen Interaction Studies: Lesson from the COVID-19 Pandemic.

COVID-19 outbreak displayed presumably an increased accumulation of SARS-CoV-2 virus during comorbid complications and a substantial variation in the mortality within and between the countries, which in turn siren us the lack of knowledge in host-pathogen interactions (HPIs). Our aim is to describe the lessons taught by the COVID-19 pandemic in the existing/missing investigations on HPI. This was from a retrospective meta-analysis of literature on "COVID-19 and comorbidity" to expose the existing paradigm gap in HPI by highlighting the omitted concepts/areas of research and new approaches to consider for the development of future therapeutics. Literature on "COVID-19 and comorbidity" apparently depicted the disparity in HPI during comorbid/immune-challenged conditions, which was reflected in the poor prognosis of the disease and failed therapeutics upon clinical trials. Moreover, the entry, adherence, multiplication, and the following establishment of infection were also varied in groups with various comorbidities. This edified that the mode of interaction of an infectious agent could vary according to the immunological and health status of the host system and hence the efficiency/success rate of treatment modalities. In addition, limited number of literature on HPI upon comorbid and immune-challenged conditions of the host manifestly indicated that there is a lack of our focus/attention on consideration of the host immune/health-specific factors in HPI studies. These alert us that the development of unambiguous therapeutic approaches is needed for a better/successful treatment of novel infectious agents in the future. By understanding the immunological state exhibited in SARS-CoV-2 infection, we conclude that the COVID-19 pandemic has taught us a great lesson that our current understanding of HPI is insufficient to fight and conquer novel infections in real life. Hence, newer approaches are obligatory to understand HPI in order to combat COVID-19-like outbreaks in the future, if any, and also to design novel immunogenic/nutraceutical-based therapeutics.

Flow Cytometry Analysis of Mycobacteria and Mycobacteria-Infected Immune Cells.

Flow cytometry enables the measurement of tens of features on individual cells from complex mixtures. Flow cytometry enables high-throughput quantification of cell size, gene and protein expression. In the case of studies of host-pathogen interactions, this tool provides a facile way of identifying cells that have been successfully infected by a pathogen. Several recent technological advances have greatly improved throughput and the number of features that can be simultaneously monitored by this technique. Here, we describe common workflows to study Mycobacterium tuberculosis heterogeneity and host-M. tuberculosis interactions using flow cytometry and related technologies.

Wax moth Galleria mellonella: Blessing or blight

Wax moth is the ubiquitous pest of honeybees globally. Wax moth larva could be a terrible menace to the beekeepers as it turns the entire wax combs to a pile of debris and frass. Wax moth larva is a voracious eater tends to feed on wax combs, cast off larval bee skin, pollen, brood and honey more seriously forcing entire colony to abscond. Despite of being a nuisance to honeybees the wax moth is a well-accepted inexpensive insect model to study host pathogen interactions and various antimicrobial compounds. The unique ability of the larvae of wax moth to biodegrade plastic is one of the major breakthroughs in the history of science. The present article reviewed the taxonomical position, life cycle and control measures of wax moth in addition to its role in plastic degradation. Eventually, this review article also throws the light on both the abilities of Galleria mellonella of being a pest as well as an efficient plastic bio degrader.

Innate immunity in C. elegans.

In its natural habitat, C. elegans encounters a wide variety of microbes, including food, commensals and pathogens. To be able to survive long enough to reproduce, C. elegans has developed a complex array of responses to pathogens. These activities are coordinated on scales that range from individual organelles to the entire organism. Often, the response is triggered within cells, by detection of infection-induced damage, mainly in the intestine or epidermis. C. elegans has, however, a capacity for cell non-autonomous regulation of these responses. This frequently involves the nervous system, integrating pathogen recognition, altering host biology and governing avoidance behavior. Although there are significant differences with the immune system of mammals, some mechanisms used to limit pathogenesis show remarkable phylogenetic conservation. The past 20 years have witnessed an explosion of host-pathogen interaction studies using C. elegans as a model. This review will discuss the broad themes that have emerged and highlight areas that remain to be fully explored.

A Perspective on Organoids for Virology Research.

Animal models and cell lines are invaluable for virology research and host–pathogen interaction studies. However, it is increasingly evident that these models are not sufficient to fully understand human viral diseases. With the advent of three-dimensional organotypic cultures, it is now possible to study viral infections in the human context. This perspective explores the potential of these organotypic cultures, also known as organoids, for virology research, antiviral testing, and shaping the virology landscape.

Opportunities and Challenges in Studies of Host-Pathogen Interactions and Management of Verticillium dahliae in Tomatoes.

Tomatoes (Solanum lycopersicum L.) are a valuable horticultural crop that are grown and consumed worldwide. Optimal production is hindered by several factors, among which Verticillium dahliae, the cause of Verticillium wilt, is considered a major biological constraint in temperate production regions. V. dahliae is difficult to mitigate because it is a vascular pathogen, has a broad host range and worldwide distribution, and can persist in soil for years. Understanding pathogen virulence and genetic diversity, host resistance, and plant-pathogen interactions could ultimately inform the development of integrated strategies to manage the disease. In recent years, considerable research has focused on providing new insights into these processes, as well as the development and integration of environment-friendly management approaches. Here, we discuss the current knowledge on the race and population structure of V. dahliae, including pathogenicity factors, host genes, proteins, enzymes involved in defense, and the emergent management strategies and future research directions for managing Verticillium wilt in tomatoes.

Corneal Infection Models: Tools to Investigate the Role of Biofilms in Bacterial Keratitis.

Bacterial keratitis is a corneal infection which may cause visual impairment or even loss of the infected eye. It remains a major cause of blindness in the developing world. Staphylococcus aureus and Pseudomonas aeruginosa are common causative agents and these bacterial species are known to colonise the corneal surface as biofilm populations. Biofilms are complex bacterial communities encased in an extracellular polymeric matrix and are notoriously difficult to eradicate once established. Biofilm bacteria exhibit different phenotypic characteristics from their planktonic counterparts, including an increased resistance to antibiotics and the host immune response. Therefore, understanding the role of biofilms will be essential in the development of new ophthalmic antimicrobials. A brief overview of biofilm-specific resistance mechanisms is provided, but this is a highly multifactorial and rapidly expanding field that warrants further research. Progression in this field is dependent on the development of suitable biofilm models that acknowledge the complexity of the ocular environment. Abiotic models of biofilm formation (where biofilms are studied on non-living surfaces) currently dominate the literature, but co-culture infection models are beginning to emerge. In vitro, ex vivo and in vivo corneal infection models have now been reported which use a variety of different experimental techniques and animal models. In this review, we will discuss existing corneal infection models and their application in the study of biofilms and host-pathogen interactions at the corneal surface.

Severity assessment in the Nicotiana tabacum-Xylella fastidiosa subsp. pauca pathosystem: design and interlaboratory validation of a standard area diagram set

Though an experimental host of Xylella fastidiosa, tobacco (Nicotiana tabacum) is an excellent plant model for biological and functional genomics studies of host-pathogen interactions involving X. fastidiosa. Symptoms induced by X. fastidiosa subsp. pauca on tobacco have been characterized, but severity assessment relies on an ordinal scale developed for symptoms on citrus. We designed a standard area diagram set (SADs) to aid in the visual estimation of percent area affected (% severity) and performed a multi-laboratory validation on tobacco cv. Havana, inoculated with X. fastidiosa subsp. pauca. Inoculated plants were monitored over time and digital images of the symptoms recorded. Three different software programs (APS Asses, ImageJ, and Leaf Doctor) were used to segment the images and calculate percent severity. Ten true-color images composed a 10-image SADs (0.5, 5, 10, 15, 25, 35, 45, 55, 65, and 75%). Fifty raters at four research groups (laboratories) assigned percent severity first without and then with the SADs on the same testing image set of 40 images following the same instructions provided by an examiner at each laboratory. The means of percent severity by all software programs were assumed to represent the actual percent severity given the perfect agreement between them. The unaided estimates were less precise, biased towards overestimation (up to 50% points) and less concordant among raters. Accuracy and precision varied considerably among the raters, and the effect of the SADs on the overall concordance to the actual severity was dependent on the laboratory; in all but one laboratory, the group means of the agreement index statistically improved. The between-rater agreement improved in all laboratories when using the SADs. The SADs may help to standardize and allow comparison across different laboratories, but care should be taken during instructions on how to discriminate symptoms that are not readily discernible as well as on how to correctly use the SADs.

Enterobactin Deficiency in a Coliform Mastitis Isolate Decreases Its Fitness in a Murine Model: A Preliminary Host-Pathogen Interaction Study.

Iron is an essential nutrient for bacterial growth. Therefore, bacteria have evolved chelation mechanisms to acquire iron for their survival. Enterobactin, a chelator with high affinity for ferric iron, is secreted by Escherichia coli and contributes to its improved bacterial fitness. In this preliminary study, we evaluated enterobactin deficiency both in vitro and in vivo in the context of E. coli mastitis. Firstly, we showed that expression of lipocalin 2, a protein produced by the host that is able to both bind and deplete enterobactin, is increased upon E. coli infection in the cow's mastitic mammary gland. Secondly, we demonstrated in vitro that enterobactin deficiency does not alter interleukin (IL)-8 expression in bovine mammary epithelial cells and its associated neutrophil recruitment. However, a significantly increased reactive oxygen species production of these neutrophils was observed. Thirdly, we showed there was no significant difference in bacterial in vitro growth between the enterobactin-deficient mutant and its wild-type counterpart. However, when further explored in a murine model for bovine mastitis, the enterobactin-deficient mutant vs. the wild-type strain revealed a significant reduction of the bacterial load and, consequently, a decrease in pro-inflammatory cytokines (IL-1α,−1β,−4,−6, and−8). A reduced neutrophilic influx was also observed immunohistochemically. These findings therefore identify interference of the enterobactin iron-scavenging mechanism as a potential measure to decrease the fitness of E. coli in the mastitic mammary gland.

Histo-blood group antigens of glycosphingolipids predict susceptibility of human intestinal enteroids to norovirus infection

The molecular mechanisms behind infection and propagation of human restricted pathogens such as human norovirus (HuNoV) have defied interrogation because they were previously unculturable. However, human intestinal enteroids (HIEs) have emerged to offer unique ex vivo models for targeted studies of intestinal biology, including inflammatory and infectious diseases. Carbohydrate-dependent histo-blood group antigens (HBGAs) are known to be critical for clinical infection. To explore whether HBGAs of glycosphingolipids contribute to HuNoV infection, we obtained HIE cultures established from stem cells isolated from jejunal biopsies of six individuals with different ABO, Lewis, and secretor genotypes. We analyzed their glycerolipid and sphingolipid compositions and quantified interaction kinetics and the affinity of HuNoV virus-like particles (VLPs) to lipid vesicles produced from the individual HIE-lipid extracts. All HIEs had a similar lipid and glycerolipid composition. Sphingolipids included HBGA-related type 1 chain glycosphingolipids (GSLs), with HBGA epitopes corresponding to the geno- and phenotypes of the different HIEs. As revealed by single-particle interaction studies of Sydney GII.4 VLPs with glycosphingolipid-containing HIE membranes, both binding kinetics and affinities explain the patterns of susceptibility toward GII.4 infection for individual HIEs. This is the first time norovirus VLPs have been shown to interact specifically with secretor gene-dependent GSLs embedded in lipid membranes of HIEs that propagate GII.4 HuNoV ex vivo, highlighting the potential of HIEs for advanced future studies of intestinal glycobiology and host-pathogen interactions.

Disease sniffing robots to apps fixing plant diseases: applications of artificial intelligence in plant pathology—a mini review

The recent advancements in artificial intelligence (AI) and machine learning have wide applications in plant pathology with sensors, drones, robots and intelligent monitoring systems. Computer vision based phenotyping of plant stress, diagnostics and severity assessment of plant diseases has gained momentum in horticultural and field crops. Internet of things based on networking sensors for biomarkers of disease like volatile organic compounds are being used for early detection and prediction of plant diseases and host-pathogen interaction studies. Unmanned arial vehicles are employed for phenotyping orchards for precise application of plant protection chemicals. Smartphone based field diagnostics are gaining popularity across the world especially in the remote locations where the laboratory diagnostics of diseases is difficult. AI in plant pathology is still at its infancy. Integration of AI and augmented reality will enhance the accuracy and automation for remote diagnostics of plant diseases and precision plant protection. The “self sufficient, disease free, perfect plant” concept will soon become reality with the help of plant-robot bio-hybrids. This mini review explores the present status of AI technologies in plant pathology and tries to find answers for the questions like what are the common platforms used, which are the diseases where technology is applied, what are the challenges and the future prospects.

Nematode-Encoded RALF Peptide Mimics Facilitate Parasitism of Plants through the FERONIA Receptor Kinase

The molecular mechanism by which plants defend against plant root-knot nematodes (RKNs) is largely unknown. The plant receptor kinase FERONIA and its peptide ligands, rapid alkalinization factors (RALFs), regulate plant immune responses and cell expansion, which are two important factors for successful RKN parasitism. In this study, we found that mutation of FERONIA in Arabidopsis thaliana resulted in plants showing low susceptibility to the RKN Meloidogyne incognita. To identify the underlying mechanisms associated with this phenomenon, we identified 18 novel RALF-likes from multiple species of RKNs and showed that two RALF-likes (i.e., MiRALF1 and MiRALF3) from M. incognita were expressed in the esophageal gland with high expression during the parasitic stages of nematode development. These nematode RALF-likes also possess the typical activities of plant RALFs and can directly bind to the extracellular domain of FERONIA to modulate specific steps of nematode parasitism-related immune responses and cell expansion. Genetically, both MiRALF1/3 and FERONIA are required for RKN parasitism in Arabidopsis and rice. Collectively, our study suggests that nematode-encoded RALFs facilitate parasitism via plant-encoded FERONIA and provides a novel paradigm for studying host–pathogen interactions.

Genome-Scale Metabolic Model of Xanthomonas phaseoli pv. manihotis: An Approach to Elucidate Pathogenicity at the Metabolic Level.

Xanthomonas phaseoli pv. manihotis (Xpm) is the causal agent of cassava bacterial blight, the most important bacterial disease in this crop. There is a paucity of knowledge about the metabolism of Xanthomonas and its relevance in the pathogenic process, with the exception of the elucidation of the xanthan biosynthesis route. Here we report the reconstruction of the genome-scale model of Xpm metabolism and the insights it provides into plant–pathogen interactions. The model, iXpm1556, displayed 1,556 reactions, 1,527 compounds, and 890 genes. Metabolic maps of central amino acid and carbohydrate metabolism, as well as xanthan biosynthesis of Xpm, were reconstructed using Escher (https://escher.github.io/) to guide the curation process and for further analyses. The model was constrained using the RNA-seq data of a mutant of Xpm for quorum sensing (QS), and these data were used to construct context-specific models (CSMs) of the metabolism of the two strains (wild type and QS mutant). The CSMs and flux balance analysis were used to get insights into pathogenicity, xanthan biosynthesis, and QS mechanisms. Between the CSMs, 653 reactions were shared; unique reactions belong to purine, pyrimidine, and amino acid metabolism. Alternative objective functions were used to demonstrate a trade-off between xanthan biosynthesis and growth and the re-allocation of resources in the process of biosynthesis. Important features altered by QS included carbohydrate metabolism, NAD(P)+ balance, and fatty acid elongation. In this work, we modeled the xanthan biosynthesis and the QS process and their impact on the metabolism of the bacterium. This model will be useful for researchers studying host–pathogen interactions and will provide insights into the mechanisms of infection used by this and other Xanthomonas species.

Identification and characterization of Dicer-like genes in leaf rust pathogen (Puccinia triticina) of wheat.

Puccinia triticina (P. triticina) is one of the most devastating fungal pathogens of wheat which causes significant annual yield loss to the crop. Understanding the gene regulatory mechanism of the biotrophic pathogen is one of the important aspects of host-pathogen interaction studies. Dicer-like genes are consideredas important mediators of RNAi-based gene regulation. In this study, we report the presence of three Dicer-like genes (Pt-DCL1, Pt-DCL2, Pt-DCL3) in P. triticina genome identified through computational and biological analyses. Quantitative real-time PCR studies revealed an increase in the expression of these genes in germinating spore stages. Heterologous expression combined with mass spectrometry analysis of Pt-DCL2 confirmed the presence of a canonical Dicer-like gene in P. triticina. Phylogenetic analysis of the Pt-DCLs with the Dicer-like proteins from other organisms showed a distinct cluster of rust pathogens from the order Pucciniales. The results indicated a species-specific duplication of Dicer-like genes within the wheat rust pathogens. This study, for the first time, reports the presence of Dicer-dependent RNAi pathway in P. triticina that may play a role in gene regulatory mechanism of the pathogen during its development. Our study serves as a vital source of information for further RNAi-based molecular studies for better understanding and management of the wheat leaf rust disease.

Time-resolved RNA-seq provided a new understanding of intestinal immune response of European eel (Anguilla anguilla) following infection with Aeromonas hydrophila

No studies systematically examined the intestinal immune response for yellow stage of European eel (Anguilla anguilla) with Aeromonas hydrophila infection by time-resolved RNA-seq. Here, we examined transcriptional profiles of the intestines at three-time points following infection with A. hydrophila. Intraperitoneal injections caused mortalities within 48 h post-injection (hpi), with the survival rate 87.5% at 24 hpi and 83.9% at 48 hpi. The result from KEGG pathway enrichment analysis showed that the immune related “cytosolic DNA-sensing pathway” was significantly enriched at the first and second time points (6 hpi and 18 hpi), with the up-regulated expression of irf3, il1b, tnfaip3, cxcl8a, ap1-2, c-fos, polr3d, polr3g and polr3k both at 6 hpi and 18 hpi, but not at the third time point (36 hpi). According to the KEGG annotation, 326 immune and inflammation-related DEGs were found. The co-expression network of those 326 DEGs revealed the existence of three modules, and tlr1 was found to be in the center of the biggest module which contained massive DEGs from “signal transduction” and “transport and catabolism”. The c3 isoforms showed different expression pattern among the three time points, indicating a unique activation of complement systems at 18 hpi. Furthermore, two cathelicidins (aaCATH_1 and aaCATH_2) were highly up-regulated at the first two time points, and the bacterial growth inhibition assay revealed their antibacterial properties against A. hydrophila. Our data indicated the important roles of cytosolic DNA-sensing pathway, as well as transcripts including tlr1, c3, polr and cathelicidins in the intestine of A. anguilla in response to A. hydrophila infection. The present study will provide leads for functional studies of host-pathogen interactions.