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Introduction: Bacterial resistance to multiple drugs is increasing at an alarming rate in current era and nanotechnology is one of the effective and novel approaches to overcome drug resistance. Methods: Zinc Oxide Nanoparticles (ZnO NPs) has stronger antibacterial activity and is regarded as bio-safe nanomaterial. The aim of present study is to synthesize the ZnO NPs using Aloe vera leaves extract and to investigate the synergistic effects and antioxidant actions of bio-synthesized ZnO NPs against gram negative bacteria E.coli and K. pneumoniae. The synergistic effect of β-lactam antibiotics (meropenem and ciprofloxacin) was tested along with ZnO NPs by using Kirby’s disc diffusion assay. The antioxidant activity was investigated by α, α-diphenyl-β-picrylhydrazyl (DPPH) method. Results: Results revealed that the antibacterial activity of the selected antibiotics was much enhanced by ZnO NPs than the antibiotics alone. The resistant antibiotic (ciprofloxacin) became sensitive when combined with ZnO NPs. The antioxidant activity reveals that biosynthesized ZnO NPs possess significantly higher (p Conclusion: The findings reveal that biosynthesized ZnO NPs have much more eco-friendly approach. It can act as a strong potentiator of β-lactam antibiotics and put forward the possibility to use them effectively in targeted drug delivery, pharmaceuticals and biomedical fields.

Labeo rohita was challenged with Fusarium oxysporum to demonstrate a thorough immune response. Significant alterations in hematological parameters viz; erythrocyte, hemoglobin, hematocrit, mean corpuscular hemoglobin concentration, mean corpuscular volume, packed cell volume, platelets, and leukocytes were observed. Total serum protein, albumin, globulin, A/G ratio, aspartate aminotransferase, alkaline aminotransferase, alkaline phosphatase activity, and cholesterol were also altered in the infected fish. Marked histopathological lesions in gills, liver, and kidney (fusion of gill lamella, necrosis, edema, vacuolization, glomerulus congestion, dilated bowman's space and renal tubules alterations) were recorded. Tissue specific gene expression data showed that interleukin was upregulated in infected fish gills, liver, and kidney but was downregulated in blood. Tumor necrosis factor-α expression was upregulated in blood, liver, and kidney and was downregulated in gills. Upregulation of interferon-γ was observed in blood, gills, and kidney but downregulated in liver. All the cellular and molecular findings from this study can be taken as critical biomarkers in the advancement of diagnostic techniques against fungal infections as a part of wide-ranging programs of systematic and comprehensive monitoring of fish health and immune system.

Heat shock proteins (HSP) are highly conserved in their structure and released in case of stress. Increased metallothionein (MT) synthesis is associated with increased capacity for binding heavy metals. Healthy juveniles of grass carp were exposed to sublethal dose (1.495 mg L-1) of cadmium for 28 days. Simultaneously, a control group was also run to compare difference of total RNA expression levels in cadmium-treated and control groups. The cadmium levels in the tissues of treated fish recorded were 1.78 ± 0.10 mg L − 1 , 1.60 ± 0.04 mg L − 1 , and 2.00 ± 0.05 mg L − 1 , respectively. Several histological alterations including edema, hemorrhage, dilated sinusoids, hypertrophy, hyperplasia, congestion of central vein, and nuclear alterations were observed in cadmium-exposed fish. Stress gene (metallothionein and heat shock proteins) mRNA transcription levels were studied by mRNA extraction and cDNA preparation by using PCR. The expression level of heat shock protein gene was higher as compared to metallothionein and beta-2-microglobulin gene after cadmium exposure. This study reports various stress-related immune-responsive changes of immune proteins, heat shock proteins, metallothionein, and histopathological changes in fish due to cadmium toxicity that make the fish immunocompromised which may be considered as the biomarkers of cadmium toxicity in other experimental species.

Breast cancer is a major cause of cancer related deaths in women worldwide. Available treatments pose serious limitations such as systemic toxicity, metastasis, tumor recurrence, off-target effects, and drug resistance. In recent years, phytochemicals such as secondary metabolites due to their effective anticancer potential at very low concentration have gained attention. Aim of the study was to evaluate anticancer potential of Citrullus colocynthis and its possible molecular targets on MCF-7, a human breast cancer cell line. Methanolic extract of leaves was prepared and fractionated by solvents (n-hexane, chloroform, ethyl acetate and n-butanol) with increasing polarity. Bioassays and gene expression regulation was conducted to evaluate the anticancer activity, proliferation rate and cell cycle regulation of breast cancer cells treated with extract and its fractions, separately. Results showed a significant anticancer activity of methanolic extract of C. colocynthis and two of its fractions prepared with chloroform and ethyl acetate. Bioassays depicted significant decrease in proliferation and growth potential along with cell cycle arrest of treated cells compared to control untreated cells. Expression regulation of genes further confirmed the cell cycle arrest through significant upregulation of cyclin-CDK inhibitors (p21 and p27) and cell cycle checkpoint regulators (HUS1, RAD1, ATM) followed by downregulation of downstream cell cycle progression genes (Cyclin A, Cyclin E, CDK2). It is concluded that C. colocynthis arrests cell cycle in human breast cancer cells through expression regulation of cyclin-CDK inhibitors and with further research can be proposed for therapeutic interventions.

Nanotechnology-based strategies can be useful tools to combat the alarming challenge of antibacterial resistance. Nanoparticles can be used as alternatives or supplementary in enhancing the effectiveness of the available drugs to control bacterial proliferation. Here, we evaluated the vibrant anti-proliferative role of silver nanoparticle (AgNPs; 31 ± 5 nm) against fish pathogen Pseudomonas aeruginosa (P. aeruginosa) to inhibit the infection in Labeo rohita. Fish was divided into four groups: control untreated (Con), 10 mg/l AgNPs exposed (NPs), P. aeruginosa challenged (Inf) and combinedly treated with AgNPs and P. aeruginosa (NPs + Inf). After 14 days of treatment changes in different bio-indicators including hemoglobin hematological parameters (hemoglobin, total erythrocyte count, hematocrit, total leukocyte count), biochemical parameters (total protein, albumin, globulin), oxidative biomarkers [malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT)] and histological parameters of kidney and liver were observed. We found that infected fish (Inf) showed more symptoms of infection as compared to fish given combined treatment of AgNPs and P. aeruginosa (NPs + Inf). Aforementioned bio-indicators in AgNPs-exposed group (NPs) were comparable to control, showing that AgNPs were not toxic at this concentration. Our results thus show that AgNPs hindered the bacterial proliferation in challenged fish. This was further confirmed by CFU assays of muscle tissues collected from Inf and NPs + Inf groups. The finding of this study suggested AgNPs as potential candidates to prevent bacterial proliferation which could be beneficial for biomedical applications.

The present study aims to assess the induced nanotoxicity of arsenic nanoparticles (AsNPs) on different organs of fresh water fish Labeo rohita. AsNPs were synthesized by chemical reduction method using sodium arsenite as precursor, ice-cold sodium borohydride as reducing agent, and sodium hydroxide to adjust the solution pH. The synthesized AsNPs were characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM) for optical, structural, and morphological investigations. The UV-Vis absorption peaks occurring at around 300 nm indicated the presence of AsNPs in colloidal sample. The rhombohedral crystalline nature and metallic purity of AsNPs with crystallite size of 30 ± 1 nm were confirmed by characteristic peaks of XRD pattern. The SEM micrograph revealed the almost spherical shape and 40 ± 10 nm average size prepared AsNPs. For assessment of induced nanotoxicity, juveniles of Labeo rohita (L. rohita) were exposed to three different concentrations of AsNPs (namely, 1, 10, and 20 mg/L) for 30 days (n = 15 per group), and the control fish was kept untreated. It was observed that the routine behavior activities (such as swimming, mutual interactions, and feed intake) were affected by AsNPs. The growth of AsNPs treated fish was found retarded as compared to the control fish. Total erythrocyte count, total leukocyte count, and hemoglobin and hematocrit values were low in the AsNPs treated fish. Immunobiochemical assays revealed that protein level was altered in the AsNPs treated fish. The levels of antioxidant enzymes catalase and superoxide dismutase were low in the treated fish. The histological alteration induced by AsNPs in liver, gills, and kidneys demonstrated the damage in form of glomerulus shrinkage, vacuolation, inflammation, necrosis, lamellar disorganization, and hemorrhage in comparison with untreated fish. The results of the present study indicate that AsNPs exposure causes behavior, growth, hematology, immunobiochemical, and histological shortcomings in L. rohita.

The marvelous self-cleaning properties of lotus leaves and their superhydrophobic nature inspire the researchers to mimic such functionalities on cotton fabrics. The present work aimed to prepare and characterize true non-sticky superhydrophobic properties on cotton samples. The required combination of micro and nanostructure roughness was achieved by depositing the silica nanoparticles (SiNPs) on cotton surfaces by dip-coating method. To enhance adhesion and durability of silica particles with cotton, SiNPs were functionalized with 3-Aminopropyltriethoxysilane (APTES) before deposition. The surface energy of SiNPs coated cotton samples was lowered by functionalizing with 1H, 1H, 2H, 2H-Perfluorooctyltriethoxysilane (PFOTS). The morphology, surface chemistry and wettability characterizations of prepared samples were performed by scanning electron microscope (SEM), energy dispersive X-ray analysis (EDX) and contact angle measurement system (goniometer) respectively. SEM micrographs revealed the nice decoration of SiNPs (having particles size range 90-150 nm) on the cotton fibers leaving to the hierarchical roughness. The EDX analysis confirmed the coatings of SiNPs and PFOTS. Contact angle measurements exhibited the superhydrophobic nature of prepared surfaces by static water contact angle of 157±2° and sliding angle of less than 5. The functionalized SiNPs/PFOTS cotton fabrics can effectively be used for self-cleaning applications.

Magnetic cores loaded with metallic nanoparticles can be promising nano-carriers for successful drug delivery at infectious sites. We report fabrication, characteristic analysis and in vitro antibacterial performance of nanocomposites comprising cobalt cores (Co-cores) functionalized with a varied concentration of silver nanoparticles (AgNPs). A two-step polyol process synchronized with the transmetalation reduction method was used. Co-cores were synthesized with cobalt acetate, and decoration of AgNPs was carried out with silver acetate. The density of AgNPs was varied by changing the amount of silver content as 0.01, 0.1 and 0.2 g in the synthesis solution. Both AgNPs and Co-cores were spherical having a size range of 30–80 nm and 200 nm to more than 1 µm, respectively, as determined by scanning electron microscopy. The metallic nature and face-centred cubic crystalline phase of prepared nanocomposites were confirmed by X-ray diffraction. Biocompatibility analysis confirmed high cell viability of MCF7 at low concentrations of tested particles. The antibacterial performance of nanocomposites (Co@AgNPs) against Escherichia coli and Bacillus subtilis was found to be AgNPs density-dependent, and nanocomposites with the highest AgNPs density exhibited the maximum bactericidal efficacy. We therefore propose that Co@AgNPs as effective drug containers for various biomedical applications.

Macrophages are phagocytic cells from the innate immune system, which forms the first line of host defense against invading pathogens. These highly dynamic immune cells can adopt specific functional phenotypes, with the pro-inflammatory M1 and anti-inflammatory M2 polarization states as the two extremes. Recently, the process of macrophage polarization during inflammation has been visualized by real time imaging in larvae of the zebrafish. This model organism has also become widely used to study macrophage responses to microbial pathogens. To support the increasing use of zebrafish in macrophage biology, we set out to determine the complete transcriptome of zebrafish larval macrophages. We studied the specificity of the macrophage signature compared with other larval immune cells and the macrophage-specific expression changes upon infection. We made use of the well-established mpeg1, mpx, and lck fluorescent reporter lines to sort and sequence the transcriptome of larval macrophages, neutrophils, and lymphoid progenitor cells, respectively. Our results provide a complete dataset of genes expressed in these different immune cell types and highlight their similarities and differences. Major differences between the macrophage and neutrophil signatures were found within the families of proteinases. Furthermore, expression of genes involved in antigen presentation and processing was specifically detected in macrophages, while lymphoid progenitors showed expression of genes involved in macrophage activation. Comparison with datasets of in vitro polarized human macrophages revealed that zebrafish macrophages express a strongly homologous gene set, comprising both M1 and M2 markers. Furthermore, transcriptome analysis of low numbers of macrophages infected by the intracellular pathogen Mycobacterium marinum revealed that infected macrophages change their transcriptomic response by downregulation of M2-associated genes and overexpression of specific M1-associated genes. Among the infection-induced genes, a homolog of the human CXCL11 chemokine gene, cxcl11aa, stood out as the most strongly overexpressed M1 marker. Upregulation of cxcl11aa in Mycobacterium-infected macrophages was found to require the function of Myd88, a critical adaptor molecule in the Toll-like and interleukin 1 receptor pathways that are central to pathogen recognition and activation of the innate immune response. Altogether, our data provide a valuable data mining resource to support infection and inflammation research in the zebrafish model.

Macrophages are phagocytic cells from the innate immune system, which forms the first line of host defense against invading pathogens. These highly dynamic immune cells can adopt specific functional phenotypes, with the pro-inflammatory M1 and anti-inflammatory M2 polarization states as the two extremes. Recently, the process of macrophage polarization during inflammation has been visualized by real time imaging in larvae of the zebrafish. This model organism has also become widely used to study macrophage responses to microbial pathogens. To support the increasing use of zebrafish in macrophage biology, we set out to determine the complete transcriptome of zebrafish larval macrophages. We studied the specificity of the macrophage signature compared with other larval immune cells and the macrophage-specific expression changes upon infection. We made use of the well-established mpeg1, mpx, and lck fluorescent reporter lines to sort and sequence the transcriptome of larval macrophages, neutrophils, and lymphoid progenitor cells, respectively. Our results provide a complete dataset of genes expressed in these different immune cell types and highlight their similarities and differences. Major differences between the macrophage and neutrophil signatures were found within the families of proteinases. Furthermore, expression of genes involved in antigen presentation and processing was specifically detected in macrophages, while lymphoid progenitors showed expression of genes involved in macrophage activation. Comparison with datasets of in vitro polarized human macrophages revealed that zebrafish macrophages express a strongly homologous gene set, comprising both M1 and M2 markers. Furthermore, transcriptome analysis of low numbers of macrophages infected by the intracellular pathogen Mycobacterium marinum revealed that infected macrophages change their transcriptomic response by downregulation of M2-associated genes and overexpression of specific M1-associated genes. Among the infection-induced genes, a homolog of the human CXCL11 chemokine gene, cxcl11aa, stood out as the most strongly overexpressed M1 marker. Upregulation of cxcl11aa in Mycobacterium-infected macrophages was found to require the function of Myd88, a critical adaptor molecule in the Toll-like and interleukin 1 receptor pathways that are central to pathogen recognition and activation of the innate immune response. Altogether, our data provide a valuable data mining resource to support infection and inflammation research in the zebrafish model.

The current study aimed to evaluate lipid profile parameters in patients with chronic kidney disease (CKD). This case-control study was conducted at the Zoology Department of Lahore College for Women University, Lahore, Pakistan, from September 2016 to June 2017 and analysed lipid profiles of patients with CKD attending different hospitals of Lahore. Of the randomly collected 100 blood samples, 50 were healthy, while the remaining 50 were patients with CKD. Lipid profile parameters were analysed using Hitachi 704 Analyser. For data analysis Chi-square test was used by means of SSPS 17. Results showed that all the parameters, including high density lipoprotein, triglycerides and low density lipoprotein were considerably altered in patients with CKD as compared to the control group. The values of triglycerides and LDL parameters were higher, while HDL cholesterol was lower in patients with CKD as compared to controls.

The fabrication, characterization, and CO2 gas detection performance of single component-based and hetero-nanostructure-based optical gas sensors are reported in the present work. Single component-based structures include (i) TiO2 thin films with varied film thickness (37.45 nm, 51.92 nm, and 99.55 nm) fabricated via the RF sputtering system for different deposition times and (ii) silver nanoparticles (AgNPs) deposited on the glass substrate by the wet chemical method. Hetero-nanostructures were achieved by decorating the AgNPs on the predeposited TiO2 thin films. The structural, morphological, and optical characteristics of prepared samples were studied by X-ray diffraction (XRD), scanning electron microscopy (SEM), and ellipsometry, respectively. XRD analysis of AgNPs confirmed the crystalline nature of prepared particles with average crystallite size of 21 nm, however, in the case of TiO2 films XRD results suggested amorphous structure of all as-deposited films. size 21 nm. The SEM micrographs confirmed the deposition of AgNPs on the TiO2 thin films. With increasing sputtering time, TiO2 films were found to be denser and more compact, indicating a reduced porosity and higher film thickness. CO2 gas-sensing properties were investigated by measuring the optical transmission spectra in alone air and in CO2 gaseous atmosphere at room temperature. It was observed that neither TiO2 thin films even with higher thickness nor alone AgNPs could demonstrate any substantial gas-sensing activity. Nevertheless, TiO2/AgNP hetero-nanostructured substrates exhibited excellent CO2 gas-sensing performance as indicated by a huge change in the transmission spectra. The enhanced sensing efficiency of TiO2/AgNP nanostructures owing to synergistic effects suggests a promising role of our manufactured sensors in practical applications.

Silver nanoparticles (AgNPs) of different shapes and sizes were prepared by solution-based chemical reduction routes. Silver nitrate was used as a precursor, tri-sodium citrate (TSC) and sodium borohydride as reducing agents, while polyvinylpyrrolidone (PVP) was used as a stabilizing agent. The morphology, size, and structural properties of obtained nanoparticles were characterized by scanning electron microscopy (SEM), UV-visible spectroscopy (UV-VIS), and X-ray diffraction (XRD) techniques. Spherical AgNPs, as depicted by SEM, were found to have diameters in the range of 15 to 90 nm while lengths of the edges of the triangular particles were about 150 nm. The characteristic surface plasmon resonance (SPR) peaks of different spherical silver colloids occurring in the wavelength range of 397 to 504 nm, whereas triangular particles showed two peaks, first at 392 nm and second at 789 nm as measured by UV-VIS. The XRD spectra of the prepared samples indicated the face-centered cubic crystalline structure of metallic AgNPs. The in vitro antibacterial properties of all synthesized AgNPs against two types of Gram-negative bacteria, Pseudomonas aeruginosa and Escherichia coli were examined by Kirby–Bauer disk diffusion susceptibility method. It was noticed that the smallest-sized spherical AgNPs demonstrated a better antibacterial activity against both bacterial strains as compared to the triangular and larger spherical shaped AgNPs.

The zebrafish (Danio rerio) is increasingly used as a model for studying infectious diseases. This nonmammalian vertebrate host, which is transparent at the early life stages, is especially attractive for live imaging of interactions between pathogens and host cells. A number of useful fluorescent reporter lines have recently been developed and significant advances in RNA sequencing technology have been made, which now make it possible to apply the zebrafish model for investigating changes in transcriptional activity of specific immune cell types during the course of an infection process.Here we describe how to sequence RNA extracted from fluorescently labeled macrophages obtained by cell-sorting of 5-day-old zebrafish larvae of the transgenic Tg(mpeg1:Gal4-VP16);Tg(UAS-E1b:Kaede) line. This technique showed reproducible results and allowed to detect specific expression of macrophage markers in the mpeg1 positive cell population, whereas no markers specific for neutrophils or lymphoid cells were detected. This protocol has been also successfully extended to other immune cell types as well as cells infected by Mycobacterium marinum.

The zebrafish (Danio rerio) is increasingly used as a model for studying infectious diseases. This nonmammalian vertebrate host, which is transparent at the early life stages, is especially attractive for live imaging of interactions between pathogens and host cells. A number of useful fluorescent reporter lines have recently been developed and significant advances in RNA sequencing technology have been made, which now make it possible to apply the zebrafish model for investigating changes in transcriptional activity of specific immune cell types during the course of an infection process.Here we describe how to sequence RNA extracted from fluorescently labeled macrophages obtained by cell-sorting of 5-day-old zebrafish larvae of the transgenic Tg(mpeg1:Gal4-VP16);Tg(UAS-E1b:Kaede) line. This technique showed reproducible results and allowed to detect specific expression of macrophage markers in the mpeg1 positive cell population, whereas no markers specific for neutrophils or lymphoid cells were detected. This protocol has been also successfully extended to other immune cell types as well as cells infected by Mycobacterium marinum.

Zebrafish model systems for infectious disease are increasingly used for the functional analysis of molecular pattern recognition processes. These studies benefit from the high conservation level of all innate immune factors in vertebrates. Zebrafish studies are strategically well positioned for this because of the ease of comparisons with studies in other fish species of which the immune system also has been intensively studied, but that are currently still less amendable to detailed genetic or microscopic studies. In this paper we focus on Toll-like receptor (TLR) signalling factors, which currently are the best characterized in mammalian systems. We review the knowledge on TLR signalling in the context of recent advances in zebrafish studies and discuss possibilities for future approaches that can complement studies in cell cultures and rodent models. A focus in these comparisons is the role of negative control mechanisms in immune responses that appear very important in a whole organism to keep adverse systemic responses in check. We also pay much attention to comparisons with studies in common carp that is highly related to zebrafish and that because of its large body mass can complement immune studies in zebrafish.

Deficiency in Src homology region 2 domain-containing phosphatase 1/protein tyrosine phosphatase nonreceptor type 6 (SHP1/PTPN6) is linked with chronic inflammatory diseases and hematological malignancies in humans. In this study, we exploited the embryonic and larval stages of zebrafish (Danio rerio) as an animal model to study ptpn6 function in the sole context of innate immunity. We show that ptpn6 knockdown induces a spontaneous inflammation-associated phenotype at the late larval stage. Surprisingly, glucocorticoid treatment did not suppress inflammation under ptpn6 knockdown conditions but further enhanced leukocyte infiltration and proinflammatory gene expression. Experiments in a germ-free environment showed that the late larval phenotype was microbe independent. When ptpn6 knockdown embryos were challenged with Salmonella typhimurium or Mycobacterium marinum at earlier stages of development, the innate immune system was hyperactivated to a contraproductive level that impaired the control of these pathogenic bacteria. Transcriptome analysis demonstrated that Kyoto Encyclopedia of Genes and Genomes pathways related to pathogen recognition and cytokine signaling were significantly enriched under these conditions, suggesting that ptpn6 functions as a negative regulator that imposes a tight control over the level of innate immune response activation during infection. In contrast to the hyperinduction of proinflammatory cytokine genes under ptpn6 knockdown conditions, anti-inflammatory il10 expression was not hyperinduced. These results support that ptpn6 has a crucial regulatory function in preventing host-detrimental effects of inflammation and is essential for a successful defense mechanism against invading microbes.

Background MicroRNAs (miRNAs) have recently been shown to play important roles in development of the immune system and in fine-tuning of immune responses. Human miR-146 family members are known as inflammation-inducible miRNAs involved in negative feedback regulation of Toll-like receptor (TLR) signalling. Dysregulation of the miR-146 family has often been linked to inflammatory diseases and malignancies. This study reports on miR-146a and miR-146b as infection-inducible miRNAs in zebrafish, which has emerged as a model species for human disease. Results Using a custom-designed microarray platform for miRNA expression we found that both members of the zebrafish miR-146 family, miR-146a and miR-146b, were commonly induced by infection of zebrafish embryos with Salmonella typhimurium and by infection of adult fish with Mycobacterium marinum. The induction of these miRNAs was confirmed by Taqman miRNA assays. Subsequently, we used zebrafish embryos, in which adaptive immunity is not yet active, as an in vivo system to investigate the role of miR-146 in the innate immune response to S. typhimurium infection. Knockdown of traf6 and use of myd88 mutants demonstrated that the induction of miR-146a and miR-146b by S. typhimurium infection was affected by disruption of the MyD88-Traf6 pathway that mediates transduction of TLR signals and cytokine responses. In turn, knockdown of miR-146 itself had no major effects on the expression of known targets of MyD88-Traf6 signalling. Instead, RNA sequencing analysis showed that miR-146 knockdown led to an increased induction of six members of the apolipoprotein gene family in S. typhimurium-infected embryos. Conclusion Based on microarray analysis and Taqman miRNA assays we conclude that members of the miR-146 family, which is highly conserved between fish and human, are induced by bacterial infection in zebrafish in a MyD88 and Traf6 dependent manner. The combined knockdown of miR-146a and miR-146b in zebrafish embryos infected with S. typhimurium had no major effect on the expression of pro-inflammatory genes and transcription factors known to be downstream of the MyD88-Traf6 pathway. In contrast, apolipoprotein-mediated lipid transport emerged as an infection-inducible pathway under miR-146 knockdown conditions, suggesting a possible function of miR-146 in regulating lipid metabolism during inflammation.

The major cell types of the innate immune system, macrophages and neutrophils, develop during the first two days of zebrafish embryogenesis. The interaction of these immune cells with pathogenic microbes can excellently be traced in the optically transparent zebrafish embryos. Various tools and methods have recently been developed for visualizing and isolating the zebrafish embryonic innate immune cells, for establishing infections by different micro-injection techniques, and for analyzing the host innate immune response following microbial recognition. Here we provide practical guidelines for the application of these methodologies and review the current state of the art in zebrafish infectious disease research.

The major cell types of the innate immune system, macrophages and neutrophils, develop during the first two days of zebrafish embryogenesis. The interaction of these immune cells with pathogenic microbes can excellently be traced in the optically transparent zebrafish embryos. Various tools and methods have recently been developed for visualizing and isolating the zebrafish embryonic innate immune cells, for establishing infections by different micro-injection techniques, and for analyzing the host innate immune response following microbial recognition. Here we provide practical guidelines for the application of these methodologies and review the current state of the art in zebrafish infectious disease research.