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

L-asparaginase (ASNase) is the principal chemotherapeutic agent against different blood cancers. The risks associated with current clinical preparations demand screening for novel ASNases. Accordingly, the study was conducted to shortlist ASNases having clinically safer profiles from a novel niche, namely, microbes in the gut and hemolymph of apparently healthy Scylla serrata. A four-step strategic approach incorporating the essential requirements for clinically safer profiles was followed. The initial step through plate assay showed five (9.61%) potential ASNase producers. The relative prevalence of ASNase producers was higher in hemolymph (13.33%) than gut (4.5%). The positive isolates were identified as Priestia aryabhattai, Priestia megaterium, Bacillus altitudinis, Shewanella decolorationis, and Chryseomicrobium amylolyticum. Quantitative profiles revealed high ASNase production (114.29 to 287.36 U/mL) without any optimization, with an added advantage of the extracellular production. The second step for substrate specificity studies revealed the absence of L-glutaminase and urease activities in ASNases from C. amylolyticum and P. megaterium, the most desirable properties for safe clinical applications. This is the first report of glutaminase and urease-free ASNase from these two bacteria. The third step ensured type II nature of selected ASNases, the targeted form in clinical applications. The fourth step confirmed the activity and stability in human physiological conditions. Altogether, the results revealed two potential ASNases with clinically compatible profiles.

• MeSH
• asparaginasa MeSH
• Bacteria genetika   MeSH
• glutaminasa MeSH
• hemolymfa MeSH
• krabi * MeSH
• lidé MeSH
• protinádorové látky * terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The problem of a growing resistance of bacteria and other microorganisms to conventional antibiotics gave rise to a search for new potent antimicrobial agents. Insect antimicrobial peptides (AMPs) seem to be promising novel potential anti-infective therapeutics. The dipeptide β-alanyl-tyrosine (β-Ala-Tyr) is one of the endogenous insect toxins exhibiting antibacterial activity against both Gram-negative and Gram-positive bacteria. Prior to testing its other antimicrobial activities, it has to be prepared in a pure form. In this study, we have developed a capillary zone electrophoresis (CZE) method for analysis of β-Ala-Tyr isolated from the extract of the hemolymph of larvae of the fleshfly Neobellieria bullata by reversed-phase high-performance liquid chromatography (RP-HPLC). Based on our previously described correlation between CZE and free-flow zone electrophoresis (FFZE), analytical CZE separation of β-Ala-Tyr and its admixtures have been converted into preparative purification of β-Ala-Tyr by FFZE with preparative capacity of 45.5 mg per hour. The high purity degree of the β-Ala-Tyr obtained by FFZE fractionation was confirmed by its subsequent CZE analysis.

• MeSH
• antiinfekční látky chemie   izolace a purifikace   MeSH
• dipeptidy chemie   izolace a purifikace   MeSH
• elektroforéza metody   MeSH
• hemolymfa chemie   MeSH
• larva chemie   MeSH
• Sarcophagidae chemie   MeSH
• vysokoúčinná kapalinová chromatografie 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

Pesticides can enter aquatic environments potentially affecting non-target organisms. Unfortunately, the effects of such substances are still poorly understood. This study investigated the effects of the active neonicotinoid substance thiacloprid (TH) and the commercial product Calypso 480 SC (CA) (active compound 40.4% TH) on Mytilus galloprovincialis after short-term exposure to sublethal concentrations. Mussels were tested for seven days to 0, 1, 5 and 10 mg L-1 TH and 0, 10, 50 and 100 mg L-1 CA. For this purpose, several parameters, such as cell viability of haemocytes and digestive cells, biochemical haemolymph features, superoxide dismutase (SOD) and catalase (CAT) enzymatic activity of gills and digestive gland, as well as histology of such tissues were analysed. The sublethal concentrations of both substances lead to abatement or completely stopping the byssal fibres creation. Biochemical analysis of haemolymph showed significant changes (P < 0.01) in electrolytes ions (Cl-, K+, Na+, Ca2+, S-phosphor), lactate dehydrogenase (LDH) enzyme activity and glucose concentration following exposure to both substances. The TH-exposed mussels showed significant imbalance (P < 0.05) in CAT activity in digestive gland and gills. CA caused significant decrease (P < 0.05) in SOD activity in gills and in CAT activity in both tissues. Results of histological analyses showed severe damage in both digestive gland and gills in a time- and concentration-dependent manner. This study provides useful information about the acute toxicity of a neonicotinoid compound and a commercial insecticide on mussels. Nevertheless, considering that neonicotinoids are still widely used and that mussels are very important species for marine environment and human consumption, further researches are needed to better comprehend the potential risk posed by such compounds to aquatic non-target species.

• MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• hemocyty účinky léků   MeSH
• hemolymfa účinky léků   MeSH
• insekticidy toxicita   MeSH
• katalasa metabolismus   MeSH
• Mytilus účinky léků   MeSH
• neonikotinoidy toxicita   MeSH
• superoxiddismutasa metabolismus   MeSH
• testy akutní toxicity MeSH
• thiaziny toxicita   MeSH
• viabilita buněk MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• žábry účinky léků   enzymologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In insects, external chemical defences, such as reflex bleeding, have been proved to be an efficient strategy against various predators. At the same time, significant costs of reflex bleeding can be expected because bled haemolymph is lost and all valuable components included have to be renewed. Interestingly, this issue has rarely been investigated for adult insects. In this study, we examined the immune and fitness costs of repeated reflex bleeding in adults of the invasive ladybird Harmonia axyridis, investigating several haemolymph parameters. Reflex bleeding induced twice a week for three weeks resulted in a significant reduction in haemocyte concentration, total protein content, and antimicrobial activity against Micrococcus luteus, and a marginally non-significant decrease in antimicrobial activity against Escherichia coli. Repeated reflex bleeding did not result in significant body mass reduction. Interestingly, resource limitation in the form of complete food absence did not significantly interact with reflex bleeding, even though starvation itself had a strong negative effect on all haemolymph parameters investigated and individual body mass. Daily reflex bleeding did not result in decreased fecundity of young ladybirds during the first 30 days of their adult life, but the start of ladybird reproduction was delayed by about two days. Moreover, ladybirds bleeding larger amounts of haemolymph started their reproduction significantly later. Overall, our results indicate that repeated reflex bleeding weakens a ladybird's immune system and can increase their susceptibility to pathogens, but a ladybird's reproductive potential remains almost unaffected, even by very intensive reflex bleeding.

• MeSH
• brouci imunologie   fyziologie   MeSH
• Escherichia coli patogenita   MeSH
• hemolymfa MeSH
• imunitní systém * MeSH
• Micrococcus luteus patogenita   MeSH
• rozmnožování MeSH
• tělesná hmotnost MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Juvenile hormones (JHs) are sesquiterpenoids synthesized by the corpora allata (CA). They play critical roles during insect development and reproduction. The first JH was described in 1934 as a "metamorphosis inhibitory hormone" in Rhodnius prolixus by Sir Vincent B. Wigglesworth. Remarkably, in spite of the importance of R. prolixus as vectors of Chagas disease and model organisms in insect physiology, the original JH that Wigglesworth described for the kissing-bug R. prolixus remained unidentified. We employed liquid chromatography mass spectrometry to search for the JH homologs present in the hemolymph of fourth instar nymphs of R. prolixus. Wigglesworth's original JH is the JH III skipped bisepoxide (JHSB3), a homolog identified in other heteropteran species. Changes in the titer of JHSB3 were studied during the 10-day long molting cycle of 4th instar nymph, between a blood meal and the ecdysis to 5th instar. In addition we measured the changes of mRNA levels in the CA for the 13 enzymes of the JH biosynthetic pathway during the molting cycle of 4th instar. Almost 90 years after the first descriptions of the role of JH in insects, this study finally reveals that the specific JH homolog responsible for Wigglesworth's original observations is JHSB3.

• MeSH
• biologická proměna * MeSH
• corpora allata chemie   MeSH
• epoxidové sloučeniny chemie   MeSH
• hemolymfa chemie   MeSH
• kukla chemie   fyziologie   MeSH
• nymfa chemie   fyziologie   MeSH
• Rhodnius chemie   fyziologie   MeSH
• seskviterpeny chemie   MeSH
• shazování tělního pokryvu fyziologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Here we characterized the development of the trypanosomatid Blastocrithidia raabei in the dock bug Coreus marginatus using light and electron microscopy. This parasite has been previously reported to occur in the host hemolymph, which is rather typical for dixenous trypanosomatids transmitted to a plant or vertebrate with insect's saliva. In addition, C. marginatus has an unusual organization of the intestine, which makes it refractory to microbial infections: two impassable segments isolate the anterior midgut portion responsible for digestion and absorption from the posterior one containing symbiotic bacteria. Our results refuted the possibility of hemolymph infection, but revealed that the refractory nature of the host provokes very aggressive behavior of the parasite and makes its life cycle more complex, reminiscent of that in some dixenous trypanosomatids. In the pre-barrier midgut portion, the epimastigotes of B. raabei attach to the epithelium and multiply similarly to regular insect trypanosomatids. However, when facing the impassable constricted region, the parasites rampage and either fiercely break through the isolating segments or attack the intestinal epithelium in front of the barrier. The cells of the latter group pass to the basal lamina and accumulate there, causing degradation of the epitheliocytes and thus helping the epimastigotes of the former group to advance posteriorly. In the symbiont-containing post-barrier midgut segment, the parasites either attach to bacterial cells and produce cyst-like amastigotes (CLAs) or infect enterocytes. In the rectum, all epimastigotes attach either to the cuticular lining or to each other and form CLAs. We argue that in addition to the specialized life cycle B. raabei possesses functional cell enhancements important either for the successful passage through the intestinal barriers (enlarged rostrum and well-developed Golgi complex) or as food reserves (vacuoles in the posterior end).

• MeSH
• elektronová mikroskopie MeSH
• hemolymfa parazitologie   MeSH
• Heteroptera imunologie   parazitologie   MeSH
• infekce prvoky kmene Euglenozoa imunologie   parazitologie   veterinární   MeSH
• interakce hostitele a parazita fyziologie   MeSH
• odolnost vůči nemocem MeSH
• stadia vývoje fyziologie   MeSH
• střevní sliznice diagnostické zobrazování   parazitologie   ultrastruktura   MeSH
• Trypanosomatina růst a vývoj   patogenita   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Freeze tolerance, the ability to survive internal ice formation, facilitates survival of some insects in cold habitats. Low-molecular-weight cryoprotectants such as sugars, polyols and amino acids are hypothesized to facilitate freeze tolerance, but their in vivo function is poorly understood. Here, we use a combination of metabolomics and manipulative experiments in vivo and ex vivo to examine the function of multiple cryoprotectants in the spring field cricket Gryllus veletis. Cold-acclimated G. veletis are freeze-tolerant and accumulate myo-inositol, proline and trehalose in their haemolymph and fat body. Injecting freeze-tolerant crickets with proline and trehalose increases survival of freezing to lower temperatures or for longer times. Similarly, exogenous myo-inositol and trehalose increase ex vivo freezing survival of fat body cells from freeze-tolerant crickets. No cryoprotectant (alone or in combination) is sufficient to confer freeze tolerance on non-acclimated, freeze-intolerant G. veletis. Given that each cryoprotectant differentially impacts survival in the frozen state, we conclude that small cryoprotectants are not interchangeable and likely function non-colligatively in insect freeze tolerance. Our study is the first to experimentally demonstrate the importance of non-colligative cryoprotectant function for insect freeze tolerance both in vivo and ex vivo, with implications for choosing new molecules for cryopreservation.

• MeSH
• aklimatizace * MeSH
• dlouhověkost MeSH
• Gryllidae růst a vývoj   fyziologie   MeSH
• hemolymfa fyziologie   MeSH
• kryoprotektivní látky metabolismus   MeSH
• metabolomika MeSH
• nízká teplota * MeSH
• nymfa růst a vývoj   fyziologie   MeSH
• prolin metabolismus   MeSH
• trehalosa metabolismus   MeSH
• tukové těleso fyziologie   MeSH
• zmrazování MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Increasing production of energy crops in Europe, mainly maize and rapeseed, has altered patterns of pesticide use in recent decades. The long-term effects of S-metolachlor (S-M) and of its metabolite metolachlor OA (M-OA) at the environmentally relevant concentration of 4.2 μg L-1 and at 42 μg L-1 (ten-fold concentration) on marbled crayfish (Procambarus virginalis) were evaluated in a 28-day exposure and after a subsequent 28-day recovery period. Indicators assessed were behaviour; biochemical haemolymph profile; oxidative and antioxidant parameters of gill, hepatopancreas, and muscle; and histology of hepatopancreas and gill. Results showed biochemical haemolymph profile (lactate, alanine aminotransferase, aspartate aminotransferase, inorganic phosphate), lipid peroxidation in hepatopancreas, and antioxidant parameters (catalase, reduced glutathione, glutathione S-transferase) of hepatopancreas and gill of crayfish exposed to S-M and M-OA to significantly differ from controls (P < 0.01). Antioxidant biomarker levels remained different from controls after a 28-day recovery period. Differences in behaviour including speed of movement and velocity, and histopathological damage to gill and hepatopancreas were associated with S-M and M-OA exposure and persisted after 28 days in S-M- and M-OA-free water. Results suggest harmful effects of low concentrations of S-M and its metabolite M-OA on non-target organisms and provide information for assessing their effects at environmentally relevant concentrations.

• MeSH
• acetamidy chemie   metabolismus   farmakologie   MeSH
• antioxidancia metabolismus   MeSH
• chování zvířat účinky léků   MeSH
• hemolymfa metabolismus   MeSH
• hepatopankreas metabolismus   MeSH
• peroxidace lipidů účinky léků   MeSH
• severní raci účinky léků   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH