larvae development
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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
Trilobites represent a model for 'evo-devo' studies in fossil euarthropods, owing to a rare developmental trait: the biomineralization of the dorsal exoskeleton soon after hatching. Many fossilized trilobite ontogenies thus feature early stages - the protaspides - characterized by non-articulated, calcified dorsal exoskeletons. The recent discovery of a protaspid-like fossil occurring with aglaspidid remains in Middle Ordovician strata of Siberia has been interpreted as evidence for the presence of protaspides in these distant relatives of trilobites. In this contribution, we demonstrate that this Siberian protaspis actually belongs to the asaphid trilobite Isotelus (or a related taxon), a genus likely present in the same bed. We conclude that protaspid larvae still represent a developmental trait unique to Trilobita.
- MeSH
- členovci * MeSH
- larva MeSH
- zkameněliny MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Paenibacillus larvae and Melissococcus plutonius represent the most threatening bacterial diseases of honeybee (Apis mellifera)-American and European foulbrood, respectively. For efficient control of those diseases, rapid and accurate detection of the pathogens is crucial. Therefore, we developed a novel multiplex PCR method simultaneously detecting both pathogens. To design and optimize multiplex PCR reaction, four strains of P. larvae representing four ERIC genotypes I-IV (strain DSM 7030-ERIC I, DSM 25430-ERIC II, LMG 16252-ERIC III, DSM 3615-ERIC IV) were selected. Those strains were fully sequenced using long-read sequencing (Sequel I, Pacific Biosciences). For P. larvae, the multicopy insertion sequence IS256 identified in all genotypes of P. larvae was selected to provide high sensitivity. M. plutonius was detected by plasmid pMP1 sequence and the virulence verified by following detection of ETX/MTX2 toxin responsible for pore formation in the cell membrane. As an internal control, a gene encoding for major royal jelly protein 1 specific for honeybees was selected. The method was validated on 36 clinical specimens collected from the colonies suffering from American and European foulbrood in the Czech Republic. Based on the results, sensitivity of PCR was calculated to 93.75% and specificity to 100% for P. larvae diagnosed from hive debris and 100% sensitivity and specificity for honeybee workers and larval scales as well as for diseased brood infected by M. plutonius.
- MeSH
- Enterococcaceae * MeSH
- larva mikrobiologie MeSH
- multiplexová polymerázová řetězová reakce metody MeSH
- Paenibacillus larvae * genetika MeSH
- Paenibacillus * genetika MeSH
- plazmidy genetika MeSH
- transpozibilní elementy DNA MeSH
- včely genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Larvae of the three important Central American malaria vectors, Anopheles albimanus, An. vestitipennis, and An. darlingi, are found in distinctly different habitats broadly defined by hydrology and aquatic vegetation, but little is known about the actual food quality and quantity of these habitats. Polyunsaturated fatty acids (PUFA) are of special interest, because mosquitoes require 20:5ω3 (EPA), 20:4ω6 (ARA), and 22:6ω3 (DHA) and without an adequate supply of these PUFAs they are not able to complete their life cycle. We collected samples of larvae and their corresponding habitats and analyzed their fatty acid (FA) composition to reveal if there are any species-specific and habitat-specific differences in FA composition, and if habitat FA differences can be linked to differences in the mosquito FA pattern and, ultimately, mosquito performance. We also assessed how FA of wild larvae compare to the laboratory-reared larvae. Habitats were generally low in essential PUFAs and there were no significant differences among the FA composition of habitat samples. There were significant differences in FA composition of larvae. An. darlingi contained significantly higher amounts of FA, specifically a higher content of ω-6 PUFA, represented mainly by the linoleic acid (18:2ω-6). Large differences were found between field-collected and laboratory-reared An. vestitipennis larvae, especially in the content of PUFAs. The laboratory-reared larvae contained significantly more of the total FA, ω3 PUFA, and MUFA. The laboratory-reared larvae contained three to five times more essential PUFAs, EPA, and DHA. However, there were no differences in the total dry weight of the 4(th) instar larvae between the wild vs laboratory-reared larvae. Total FA in both larvae and habitats of An. albimanus and An. darlingi were positively correlated with the concentration of particulate organic carbon and nitrogen (POC, PON) in their respective habitats, but no such correlation was found for An. vestitipennis. PUFA are a good indicator of nutritional quality, although factors controlling the success of anopheline development from larval habitats are likely to be more complex and would include the presence of predators, pathogens, and toxins as interacting factors.
The infective juveniles (IJs) of entomopathogenic nematodes (EPNs) seek out host insects and release their symbiotic bacteria into their body cavity causing septicaemia, which eventually leads to host death. The interaction between EPNs and their hosts are only partially understood, in particular the host immune responses appears to involve pathways other than phagocytosis and the canonical transcriptional induction pathways. These pathways are genetically tractable and include for example clotting factors and lipid mediators. The aim of this study was to optimize the nematode infections in Drosophila melanogaster larvae, a well-studied and genetically tractable model organism. Here we show that two nematode species namely Steinernema feltiae and Heterorhabditis bacteriophora display different infectivity toward Drosophila larvae with the latter being less pathogenic. The effects of supporting media and IJ dosage on the mortality of the hosts were assessed and optimized. Using optimum conditions, a faster and efficient setup for nematode infections was developed. This newly established infection model in Drosophila larvae will be applicable in large scale screens aimed at identifying novel genes/pathways involved in innate immune responses.
- MeSH
- Drosophila melanogaster genetika parazitologie MeSH
- interakce hostitele a parazita MeSH
- larva parazitologie MeSH
- Photorhabdus fyziologie MeSH
- Rhabditida mikrobiologie fyziologie MeSH
- Xenorhabdus fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The cryopreservation techniques proposed for embryos of the fruit fly Drosophila melanogaster are not yet ready for practical use. Alternative methods for long-term storage of D. melanogaster strains, although urgently needed, do not exist. Herein, we describe a narrow interval of low temperatures under which the larvae of D. melanogaster can be stored in quiescence for up to two months. The development of larvae was arrested at the pre-wandering stage under fluctuating thermal regime (FTR), which simultaneously resulted in diminishing the accumulation of indirect chill injuries. Our physiological, metabolomic, and transcriptomic analyses revealed that compared to larvae stored at constant low temperatures, the larvae stored under FTR conditions were able to decrease the rates of depletion of energy substrates, exploited brief warm episodes of FTR for homeostatic control of metabolite levels, and more efficiently exerted protection against oxidative damage.
The thermoregulation behavior of Lucilia sericata larvae (Diptera: Calliphoridae), a necrophagous species that feeds on vertebrate cadavers, was investigated. These larvae require high heat incomes to develop, and can elevate temperatures by forming large aggregates. We hypothesized that L. sericata larvae should continue to feed at temperatures up to 38 °C, which can be reached inside larval masses. Thermal regulation behavior such as movement between a hot food spot and colder areas was also postulated. The hypotheses were tested by tracking for 1 h the activity of single, starved third instar larvae in a Petri dish containing 1 food spot (FS) that was heated to a constant temperature of 25 °C, 34 °C or 38 °C with an ambient temperature of 25 °C. The influence of previous conspecific activity in the food on larval behavior was also tested. The crops of larvae were dissected to monitor food content in the digestive systems. Based on relative crop measurements, larvae fed at all food temperatures, but temperature strongly affected larval behavior and kinematics. The total time spent by larvae in FS and the duration of each stay decreased at high FS temperature. Previous activity of conspecifics in the food slightly increased the time spent by larvae in FS and also decreased the average distance to FS. Therefore, necrophagous L. sericata larvae likely thermoregulate during normal feeding activities by adjusting to local fluctuations in temperature, particularly inside maggot masses. By maintaining a steady internal body temperature, larvae likely reduce their development time.
- MeSH
- Diptera růst a vývoj fyziologie MeSH
- larva růst a vývoj fyziologie MeSH
- stravovací zvyklosti MeSH
- termoregulace * MeSH
- vysoká teplota MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
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
Thermal quality and predation risk are considered important factors influencing habitat patch use in ectothermic prey. However, how the predator's food requirement and the prey's necessity to avoid predation interact with their respective thermoregulatory strategies remains poorly understood. The recently developed 'thermal game model' predicts that in the face of imminent predation, prey should divide their time equally among a range of thermal patches. In contrast, predators should concentrate their hunting activities towards warmer patches. In this study, we test these predictions in a laboratory setup and an artificial environment that mimics more natural conditions. In both cases, we scored thermal patch use of newt larvae (prey) and free-ranging dragonfly nymphs (predators). Similar effects were seen in both settings. The newt larvae spent less time in the warm patch if dragonfly nymphs were present. The patch use of the dragonfly nymphs did not change as a function of prey availability, even when the nymphs were starved prior to the experiment. Our behavioral observations partially corroborate predictions of the thermal game model. In line with asymmetric fitness pay-offs in predator-prey interactions (the 'life-dinner' principle), the prey's thermal strategy is more sensitive to the presence of predators than vice versa.
- MeSH
- biologické modely MeSH
- larva fyziologie MeSH
- nymfa fyziologie MeSH
- predátorské chování fyziologie MeSH
- Salamandridae růst a vývoj fyziologie MeSH
- teplota * MeSH
- vážky růst a vývoj fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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
