Introduction. The fungal pathogen Aspergillus fumigatus can induce prolonged colonization of the lungs of susceptible patients, resulting in conditions such as allergic bronchopulmonary aspergillosis and chronic pulmonary aspergillosis.Hypothesis. Analysis of the A. fumigatus secretome released during sub-lethal infection of G. mellonella larvae may give an insight into products released during prolonged human colonisation.Methodology.Galleria mellonella larvae were infected with A. fumigatus, and the metabolism of host carbohydrate and proteins and production of fungal virulence factors were analysed. Label-free qualitative proteomic analysis was performed to identify fungal proteins in larvae at 96 hours post-infection and also to identify changes in the Galleria proteome as a result of infection.Results. Infected larvae demonstrated increasing concentrations of gliotoxin and siderophore and displayed reduced amounts of haemolymph carbohydrate and protein. Fungal proteins (399) were detected by qualitative proteomic analysis in cell-free haemolymph at 96 hours and could be categorized into seven groups, including virulence (n = 25), stress response (n = 34), DNA repair and replication (n = 39), translation (n = 22), metabolism (n = 42), released intracellular (n = 28) and cellular development and cell cycle (n = 53). Analysis of the Gallerial proteome at 96 hours post-infection revealed changes in the abundance of proteins associated with immune function, metabolism, cellular structure, insect development, transcription/translation and detoxification.Conclusion. Characterizing the impact of the fungal secretome on the host may provide an insight into how A. fumigatus damages tissue and suppresses the immune response during long-term pulmonary colonization.
- MeSH
- Aspergillus fumigatus * metabolismus MeSH
- aspergilóza mikrobiologie metabolismus MeSH
- faktory virulence metabolismus MeSH
- fungální proteiny * metabolismus genetika MeSH
- hemolymfa mikrobiologie metabolismus MeSH
- larva * mikrobiologie MeSH
- můry * mikrobiologie MeSH
- proteom analýza MeSH
- proteomika MeSH
- sekretom metabolismus MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
O-methylation is an unusual sugar modification with a function that is not fully understood. Given its occurrence and recognition by lectins involved in the immune response, methylated sugars were proposed to represent a conserved pathogen-associated molecular pattern. We describe the interaction of O-methylated saccharides with two β-propeller lectins, the newly described PLL2 from the entomopathogenic bacterium Photorhabdus laumondii, and its homologue PHL from the related human pathogen Photorhabdus asymbiotica. The crystal structures of PLL2 and PHL revealed up to 10 out of 14 potential binding sites per protein subunit to be occupied with O-methylated structures. The avidity effect strengthens the interaction by 4 orders of magnitude. PLL2 and PHL also interfere with the early immune response by modulating the production of reactive oxygen species and phenoloxidase activity. Since bacteria from Photorhabdus spp. have a complex life cycle involving pathogenicity towards different hosts, the involvement of PLL2 and PHL might contribute to the pathogen overcoming insect and human immune system defences in the early stages of infection. DATABASES: Structural data are available in PDB database under the accession numbers 6RG2, 6RGG, 6RFZ, 6RG1, 6RGU, 6RGW, 6RGJ, and 6RGR.
- MeSH
- bakteriální proteiny chemie metabolismus MeSH
- cukry metabolismus MeSH
- gramnegativní bakteriální infekce imunologie metabolismus mikrobiologie MeSH
- hemocyty imunologie metabolismus MeSH
- hemolymfa imunologie metabolismus MeSH
- imunita imunologie MeSH
- imunitní systém imunologie metabolismus MeSH
- interakce hostitele a patogenu imunologie MeSH
- lektiny chemie metabolismus MeSH
- lidé MeSH
- metylace MeSH
- můry MeSH
- Photorhabdus imunologie metabolismus fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In the temperate climates of central Europe and North America, two distinct honeybee (Apis mellifera) populations are found in colonies: short-living summer bees emerge in spring and survive until summer, whereas long-living winter bees emerge in late August and overwinter. Besides the difference in their life spans, each of these populations fulfils a different role in the colonies and individual bees have distinct physiological and immunological adaptations depending on their roles. For instance, winter worker bees have higher vitellogenin levels and larger reserves of nutrients in the fat body than summer bees. The differences between the immune systems of both populations are well described at the constitutive level; however, our knowledge of its inducibility is still very limited. In this study, we focus on the response of 10-day-old honeybee workers to immune challenges triggered in vivo by injecting heat-killed bacteria, with particular focus on honeybees that emerge and live under hive conditions. Responses to bacterial injections differed between summer and winter bees. Winter bees exhibited a more intense response, including higher expression of antimicrobial genes and antimicrobial activity, as well as a significant decrease in vitellogenin gene expression and its concentration in the hemolymph. The intense immune response observed in winter honeybees may contribute to our understanding of the relationships between colony fitness and infection with pathogens, as well as its association with successful overwintering.
- MeSH
- imunita * MeSH
- roční období MeSH
- včely MeSH
- vitelogeniny * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa MeSH
- Severní Amerika MeSH
Individuals reared in captivity are exposed to distinct selection pressures and evolutionary processes causing genetic and phenotypic divergence from wild populations. Consequently, restocking with farmed individuals may represent a considerable risk for the fitness of free-living populations. Supportive breeding on a massive scale has been established in many European countries to increase hunting opportunities for the most common duck species, the mallard (Anas platyrhynchos). It has previously been shown that mallards from breeding facilities differ genetically from wild populations and there is some indication of morphological differences. Using a common-garden experiment, we tested for differences in growth parameters between free-living populations and individuals from breeding facilities during the first 20 days of post-hatching development, a critical phase for survival in free-living populations. In addition, we compared their immune function by assessing two haematological parameters, H/L ratio and immature erythrocyte frequency, and plasma complement activity. Our data show that farmed ducklings exhibit larger morphological parameters, a higher growth rates, and higher complement activity. In haematological parameters, we observed high dynamic changes in duckling ontogeny in relation to their morphological parameters. In conclusion, our data demonstrate pronounced phenotype divergence between farmed and wild mallard populations that can be genetically determined. We argue that this divergence can directly or indirectly affect fitness of farmed individuals introduced to the breeding population as well as fitness of farmed x wild hybrids.
- MeSH
- chov MeSH
- divoká zvířata růst a vývoj imunologie MeSH
- farmy MeSH
- fenotyp MeSH
- kachny růst a vývoj imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
The aim of this study was to assess the effects of T-2 toxin-contaminated feed (at concentrations of 1.0 and 1.8 mg/kg) on the rainbow trout immune system by studying non-specific cellular and humoral immune responses and its effect on red and white blood cells. Consumption of T-2 toxin at both concentrations resulted in significantly increased erythrocyte counts and a decrease in mean corpuscular volume. While a significant decrease in mean corpuscular haemoglobin was observed at both experimental concentrations, the decrease in plasma haemoglobin was only significant at the higher T-2 toxin concentration. Higher T-2 toxin concentrations resulted in a significant increase in leukocyte and lymphocyte count, while absolute phagocyte count and counts of less mature neutrophil granulocyte forms remained unchanged at both concentrations. Non-specific humoral immunity (bactericidal activity measured as complement activation) decreased significantly in both experimental groups when compared with the control. The results of this study show that T-2 toxin in feed at a concentration range of 1.0-1.8 mg/kg influences the immunological defence mechanisms of rainbow trout.Trial registration number, MSMT-3876/2014-14; date of registration, 31/1/2014.
- MeSH
- Fusarium chemie metabolismus MeSH
- hemoglobiny metabolismus MeSH
- humorální imunita účinky léků MeSH
- kontaminace potravin analýza MeSH
- krmivo pro zvířata analýza MeSH
- Oncorhynchus mykiss krev imunologie metabolismus mikrobiologie MeSH
- počet erytrocytů MeSH
- počet leukocytů MeSH
- T-2 toxin analýza metabolismus toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
This study examined how adipokinetic hormone (AKH) and adenosine affect defense responses in Drosophila melanogaster larvae infected with entomopathogenic nematodes (EPN, Steinernema carpocapsae and Heterorhabditis bacteriophora). Three loss-of-function mutant larvae were tested: Akh1, AdoR1 (adenosine receptor), and Akh1 AdoR1. Mortality decreased in all mutants post-EPN infection compared with the control (w1118). Additionally, co-application of external AKH with EPN significantly increased mortality beyond rates observed in EPN-only treatment, while also elevating carbon dioxide production, a measure of metabolism. Furthermore trehalose levels increased in both w1118 and Akh1 larvae post-EPN infection, but the latter group exhibited a lower increase and total trehalose levels. Interestingly, baseline trehalose was relatively high in untreated AdoR1 and Akh1 AdoR1 mutants, with levels remaining unaffected by infection. Infection also elevated haemolymph lipid content overall, but the different mutations did not substantially influence this change. In contrast, haemolymph protein content dropped after EPN infection in all tested groups, but this decline was more intense among Akh1. In uninfected larvae mutations decreased antioxidative capacity in Akh1 and increased in AdoR1, however, its post-infection increases were similar in all mutants, suggesting that antioxidant response in Drosophila involves mechanisms also beyond AKH and adenosine. Furthermore, AKH application in w1118 larvae significantly increased movement distance and percentage of larval activity, but reduced velocity. Mutations of Akh and AdoR did not strongly affect locomotion.
- MeSH
- adenosin metabolismus MeSH
- antibióza * MeSH
- Drosophila melanogaster růst a vývoj mikrobiologie parazitologie fyziologie MeSH
- fyziologie bakterií MeSH
- hmyzí hormony metabolismus MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty metabolismus MeSH
- larva růst a vývoj mikrobiologie parazitologie fyziologie MeSH
- oligopeptidy metabolismus MeSH
- Rhabditida fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The effect of feeding caffeine on the behaviour and neural proteome of Galleria mellonella larvae was assessed. Caffeine was administered to larvae by force feeding and the metabolites theobromine and theophylline were subsequently detected by RP-HPLC analysis. Administration of caffeine to larvae resulted in reduced movement and a reduction in the formation of pupae. The production of the muscle relaxant theophylline may contribute to the reduction in larval movement. Analysis of the changes in proteome of the brain and surrounding tissues of caffeine fed larvae revealed an increase in the abundance of immune related proteins such as immune-related Hdd1 (6.28 fold increase) and hemolin (1.68 fold increase), ATPase associated proteins such as H+ transporting ATP synthase O subunit isoform 1 (1.87 fold increase) and H+ transporting ATP synthase delta subunit (1.53 fold increase) and proteins indicative of brain trauma such as troponin T transcript variant B, partial (1.55 fold increase). Proteins involved in development and protein degradation such as SUMO-activating enzyme subunit 1 (3.08 fold decrease) and chitin deacetylase, partial (3.67 fold decrease) were decreased in abundance. The results presented here indicate that caffeine is metabolised in a similar way in G. mellonella larvae to that in mammals and results in a variety of behavioural and developmental alterations. Utilisation of insects for studying the effects of caffeine and other neuroactive compounds may offer new insights into their mode of action and reduce the need to use mammals for this type of analysis.
- MeSH
- chování zvířat účinky léků MeSH
- hmyzí proteiny metabolismus MeSH
- kofein aplikace a dávkování metabolismus MeSH
- larva účinky léků růst a vývoj MeSH
- mozek účinky léků metabolismus MeSH
- můry účinky léků růst a vývoj MeSH
- pohyb účinky léků MeSH
- proteom metabolismus MeSH
- stimulanty centrálního nervového systému aplikace a dávkování metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Photorhabdus asymbiotica is one of the three recognized species of the Photorhabdus genus, which consists of gram-negative bioluminescent bacteria belonging to the family Morganellaceae. These bacteria live in a symbiotic relationship with nematodes from the genus Heterorhabditis, together forming a complex that is highly pathogenic for insects. Unlike other Photorhabdus species, which are strictly entomopathogenic, P. asymbiotica is unique in its ability to act as an emerging human pathogen. Analysis of the P. asymbiotica genome identified a novel fucose-binding lectin designated PHL with a strong sequence similarity to the recently described P. luminescens lectin PLL. Recombinant PHL exhibited high affinity for fucosylated carbohydrates and the unusual disaccharide 3,6-O-Me2-Glcβ1-4(2,3-O-Me2)Rhaα-O-(p-C6H4)-OCH2CH2NH2 from Mycobacterium leprae. Based on its crystal structure, PHL forms a seven-bladed β-propeller assembling into a homo-dimer with an inter-subunit disulfide bridge. Investigating complexes with different ligands revealed the existence of two sets of binding sites per monomer-the first type prefers l-fucose and its derivatives, whereas the second type can bind d-galactose. Based on the sequence analysis, PHL could contain up to twelve binding sites per monomer. PHL was shown to interact with all types of red blood cells and insect haemocytes. Interestingly, PHL inhibited the production of reactive oxygen species induced by zymosan A in human blood and antimicrobial activity both in human blood, serum and insect haemolymph. Concurrently, PHL increased the constitutive level of oxidants in the blood and induced melanisation in haemolymph. Our results suggest that PHL might play a crucial role in the interaction of P. asymbiotica with both human and insect hosts.
- MeSH
- bakteriální proteiny genetika imunologie MeSH
- interakce hostitele a patogenu imunologie MeSH
- konformace proteinů MeSH
- krystalografie rentgenová MeSH
- lektiny chemie genetika imunologie MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- Photorhabdus genetika imunologie MeSH
- povrchová plasmonová rezonance MeSH
- sekvence nukleotidů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Proliferative kidney disease (PKD) is an endoparasitic disease of salmonid fish caused by Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea). This study presents a comprehensive view on PKD development in rainbow trout (Oncorhynchus mykiss) reared at an intensive fish breeding facility, with focus on mortality, pathology/histopathology, haematological findings and immune functions. Diseased and reference fish were sampled monthly and time course of natural infection was followed up from the onset of clinical signs (September 2014) to full recovery (January 2015). PKD- associated cumulative mortality was 30% with a peak value in October, while immunohistochemical testing indicated a continuous significant decrease in T. bryosalmonae numbers from September to December; with no parasites detected in January. During peak clinical infection, a significant decrease in red blood cell counts, haematocrit values, haemoglobin concentration, along with a reduction in lymphocytes and a significant phagocyte elevation corresponding with an increase in phagocyte oxidative burst were measured in comparison to control animals. Complement activity and total immunoglobulin plasma concentrations were also elevated, though only during the initial monitoring period (September). Individuals surviving PKD, recovered and were able to fully regenerate both renal structure and haematopoietic parameters to normal levels. Changes in the red blood cell parameters indicate anaemia and a decreased oxygen transportation capacity during the clinical disease phase. Together with an increased oxygen demand at higher temperatures and decreased oxygen solubility this could lead to decompensation and elevated mortality. The stimulation of immune parameters, and especially oxidative phagocytic burst, is likely to have had a strong effect on both, regeneration and elimination of the pathogenic agent.
- MeSH
- epidemický výskyt choroby veterinární MeSH
- Myxozoa fyziologie MeSH
- náchylnost k nemoci veterinární MeSH
- nemoci ledvin patologie veterinární MeSH
- nemoci ryb parazitologie MeSH
- Oncorhynchus mykiss parazitologie MeSH
- parazitární nemoci u zvířat parazitologie MeSH
- teplota MeSH
- vodní hospodářství MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
One of the key factors that determine the interaction between hosts and their parasites is the frequency of their interactions, which depends on the locomotory behavior of both parts. To address host behavior we used natural infections involving insect pathogenic nematodes and Drosophila melanogaster larvae as hosts. Using a modified version of a recently described method (FIMTrack) to assess several parameters in larger sets of animals, we initially detected specific differences in larval food searching when comparing Drosophila strains. These differences were further influenced by the presence of nematodes. Given a choice, Drosophila larvae clearly avoided nematodes irrespective of their genetic background. Our newly developed methods will be useful to test candidate genes and pathways involved in host/pathogen interactions in general and to assess specific parameters of their interaction.
- MeSH
- Drosophila melanogaster růst a vývoj parazitologie fyziologie MeSH
- interakce hostitele a parazita MeSH
- larva parazitologie fyziologie MeSH
- lokomoce MeSH
- Rhabditida fyziologie MeSH
- stravovací zvyklosti MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
