Bee health is influenced by multiple factors, including nutrition, immunity, and parasitic pressures. Since the spread of Varroa destructor, overwintering survival has significantly declined, making it one of the most serious threats to honey bee (Apis mellifera L.) populations worldwide. Natural acaricides, such as oxalic acid (OA), are widely employed for managing Varroa mites; however, their pharmacodynamics, particularly their impacts on honey bee physiology and immunity, remain insufficiently understood. We studied effects of oxalic acid on honey bee workers. The study compared three treatments: flumethrin, OA-glycerine strips (OA-G), and OA trickling (OA-T). Twelve colonies were divided into four groups, with samples collected at five time points (0, 24, 48, 72, and 192 h). Physiological changes were assessed through markers of oxidative stress, longevity, and immune parameters. Exposure to oxalic acid via glycerine strips induced a humoral immune response in adult bees. The antimicrobial activity of hemolymph and levels of antimicrobial peptides (abaecin, apidaecin, defensin, and hymenoptaecin) were elevated between 48 and 192 h after OA-G treatment compared to the control group. In contrast, these parameters were not influenced by OA-T or flumethrin treatment. These findings suggest that OA-G strips activate the honey bee's immune system, providing insights into broader implications of OA use in beekeeping. It is crucial to determine whether the activation of humoral immune systems has positive or negative effects, as well as to develop standardized and reliable treatment protocols that ensure both - health of colonies and their effectiveness in controlling Varroa mite infestations.
- Keywords
- Beekeeping, Integrated Pest management, Oxalic acid, Pharmacodynamics, Treatment, Varroa,
- MeSH
- Acaricides * pharmacology MeSH
- Antimicrobial Peptides metabolism MeSH
- Hemolymph drug effects metabolism MeSH
- Oxalic Acid * pharmacology MeSH
- Oxidative Stress drug effects MeSH
- Pyrethrins pharmacology MeSH
- Varroidae drug effects MeSH
- Bees immunology drug effects MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Acaricides * MeSH
- Antimicrobial Peptides MeSH
- flumethrin MeSH Browser
- Oxalic Acid * MeSH
- Pyrethrins MeSH
In the current study, the effects of dietary fulvic acid supplementation at levels of 0.5, 1 and 2% were examined in white-leg shrimp, Litopenaeus vannamei. A significant increase in the weight of the shrimp was observed in the group treated with 2% fulvic acid in comparison to the control group. This may have been associated with an increased digestive efficiency, with the food conversion ratio reducing from 2.4 to 1.9, and increased hepatopancreatic amylase, protease, and lipase enzyme activities. Enhanced activity of hemolymph superoxide dismutase was suggestive of an enhanced immune capacity, while hemolymph cell count increased by 16.4 and 13.6% in shrimp receiving diets supplemented with 1 and 2% fulvic acid, respectively. Additionally, the number of large granular cells increased by 37.3% and 40.8% relative to the control in these two groups. Furthermore, the lysozyme activity increased in shrimp receiving dietary supplementation of 1% and 2% fulvic acid by 16.7% and 24.7%, respectively. Phenol oxidase activity, which activates phagocytosis and encapsulation of invading pathogens, increased in all groups supplemented with fulvic acid, with the highest activity in the 1% fulvic acid group. Overall the present results suggest that fulvic acid is a promising feed additive for white-leg shrimp super-intensive culture.
- Keywords
- Fulvic acid, Growth performance, Humic substances, Immunostimulants, Pacific white shrimp,
- MeSH
- Antioxidants * metabolism pharmacology MeSH
- Benzopyrans * pharmacology MeSH
- Hemolymph metabolism drug effects MeSH
- Animal Feed analysis MeSH
- Penaeidae * drug effects immunology metabolism MeSH
- Dietary Supplements MeSH
- Digestion drug effects MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Antioxidants * MeSH
- Benzopyrans * MeSH
- fulvic acid MeSH Browser
Ecdysteroids represent a large class of polyhydroxylated steroids which, due to their anabolic properties, are marketed as dietary supplements. Some ecdysteroids also act as important hormones in arthropods, where they regulate molting, development, and reproduction and many of these insects are miniature organisms that contain submicroliter levels of circulating biofluids. Analysis of ecdysteroids is further complicated by their very low abundance, large fluctuations during development, and difficult access to a pooled sample, which is important for quantitative measurements. In this work, we propose a new method that overcomes the described difficulties and allows validated quantification of four ecdysteroids in minimal amounts of biological material. After methanolic extraction, detectability of the ecdysteroids is increased 16- to 20-fold by conversion to their 14,15-anhydrooximes. These are further purified by pipette tip solid-phase extraction on a three-layer sorbent and subjected to HPLC-MS/MS analysis. Full validation was achieved using hemolymph from larvae of the firebug Pyrrhocoris apterus as a blank matrix and by the determination of ecdysteroids in a single Drosophila larva. The lower limit of quantifications for the four target ecdysteroids (20-hydroxyecdysone, ecdysone, makisterone A, and 2-deoxyecdysone) were 0.01; 0.1; 0.05; and 0.025 pg·ml-1 (20; 200; 100; 50 fmol ml-1), respectively, with very good accuracy, precision (expressed as relative standard deviation <15%) and recoveries (96%-119.9%). The application potential of the new method was demonstrated by quantification of ecdysteroids in various biological materials including human serum.
- Keywords
- arthropods, dietary supplementation, ecdysteroid hormones, human body fluid, quantification, submilligram sample amount, ultratrace HPLC-MS analysis,
- MeSH
- Ecdysteroids * analysis blood chemistry MeSH
- Hemolymph chemistry metabolism MeSH
- Liquid Chromatography-Mass Spectrometry MeSH
- Larva MeSH
- Tandem Mass Spectrometry * methods MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Validation Study MeSH
- Names of Substances
- Ecdysteroids * 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.
- Keywords
- Aspergillus, Galleria mellonella, fungal–host interactions, gliotoxin, proteomics,
- MeSH
- Aspergillus fumigatus * metabolism MeSH
- Aspergillosis microbiology metabolism MeSH
- Virulence Factors metabolism MeSH
- Fungal Proteins * metabolism genetics MeSH
- Hemolymph microbiology metabolism MeSH
- Larva * microbiology MeSH
- Moths * microbiology MeSH
- Proteome analysis MeSH
- Proteomics MeSH
- Secretome metabolism MeSH
- Virulence MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Virulence Factors MeSH
- Fungal Proteins * MeSH
- Proteome MeSH
Ticks are blood-feeding arachnids that are known to transmit various pathogenic microorganisms to their hosts. During blood feeding, ticks activate their metabolism and immune system to efficiently utilise nutrients from the host's blood and complete the feeding process. In contrast to insects, in which the fat body is known to be a central organ that controls essential metabolic processes and immune defense mechanisms, the function of the fat body in tick physiology is still relatively unexplored. To fill this gap, we sought to uncover the repertoire of genes expressed in the fat body associated with trachea (FB/Tr) by analyzing the transcriptome of individual, partially fed (previtellogenic) Ixodes ricinus females. The resulting catalog of individual mRNA sequences reveals a broad repertoire of transcripts encoding proteins involved in nutrient storage and distribution, as well as components of the tick immune system. To gain a detailed insight into the secretory products of FB/Tr specifically involved in inter-tissue transport and humoral immunity, the transcriptomic data were complemented with the proteome of soluble proteins in the hemolymph of partially fed female ticks. Among these proteins, the hemolipoglyco-carrier proteins were predominant. When comparing immune peptides and proteins from the fat body with those produced by hemocytes, we found that the fat body serves as a unique producer of certain immune components. Finally, time-resolved transcriptional regulation of selected immune transcripts from the FB/Tr was examined in response to experimental challenges with model microbes and analyzed by RT-qPCR. Overall, our data show that the fat body of ticks, similar to insects, is an important metabolic tissue that also plays a remarkable role in immune defense against invading microbes. These findings improve our understanding of tick biology and its impact on the transmission of tick-borne pathogens.
- MeSH
- Hemolymph * MeSH
- Ixodes * genetics metabolism MeSH
- Arthropod Proteins genetics metabolism MeSH
- Proteomics MeSH
- Gene Expression Profiling MeSH
- Fat Body metabolism MeSH
- Animals MeSH
- Check Tag
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Arthropod Proteins MeSH
Hemolymph is the circulatory fluid that fills the body cavity of crustaceans, analogous to blood in vertebrates. Hemolymph coagulation, similar to blood clotting in vertebrates, plays a crucial role in wound healing and innate immune responses. Despite extensive studies on the clotting process in crustaceans, no comparative quantitative analysis of the protein composition of non-clotted and clotted hemolymph in any decapod has been reported. In this study, we used label-free protein quantification with high-resolution mass spectrometry to identify the proteomic profile of hemolymph in crayfish and quantify significant changes in protein abundances between non-clotted and clotted hemolymph. Our analysis identified a total of two-hundred and nineteen proteins in both hemolymph groups. Furthermore, we discussed the potential functions of the top most high and low-abundant proteins in hemolymph proteomic profile. The quantity of most of the proteins was not significantly changed during coagulation between non-clotted and clotted hemolymph, which may indicate that clotting proteins are likely pre-synthesized, allowing for a swift coagulation response to injury. Four proteins still showed abundance differences (p < 0.05, fold change>2), including C-type lectin domain-containing proteins, Laminin A chain, Tropomyosin, and Reverse transcriptase domain-containing proteins. While the first three proteins were down-regulated, the last one was up-regulated. The down-regulation of structural and cytoskeletal proteins may affect the process of hemocyte degranulation needed for coagulation, while the up-regulation of an immune-related protein might be attributed to the phagocytosis ability of viable hemocytes during coagulation.
- Keywords
- Clot proteomics, Decapods, Innate immunity, Protein,
- MeSH
- Hemocytes MeSH
- Blood Coagulation physiology MeSH
- Hemolymph * metabolism MeSH
- Blood Coagulation Factors metabolism MeSH
- Proteomics MeSH
- Astacoidea * physiology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Blood Coagulation Factors 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
- Asparaginase MeSH
- Bacteria genetics MeSH
- Glutaminase MeSH
- Hemolymph MeSH
- Brachyura * MeSH
- Humans MeSH
- Antineoplastic Agents * therapeutic use MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Asparaginase MeSH
- Glutaminase MeSH
- Antineoplastic Agents * MeSH
Worker honey bees are subject to biochemical and physiological changes throughout the year. This study aimed to provide the reasons behind these fluctuations. The markers analysed included lipid, carbohydrate, and protein levels in the haemolymph; the activity of digestive enzymes in the midgut; the levels of adipokinetic hormone (AKH) in the bee central nervous system; the levels of vitellogenins in the bee venom and haemolymph; and the levels of melittin in the venom. The levels of all the main nutrients in the haemolymph peaked mostly within the period of maximal bee activity, whereas the activity of digestive enzymes mostly showed a two-peak course. Furthermore, the levels of AKHs fluctuated throughout the year, with modest but significant variations. These data suggest that the role of AKHs in bee energy metabolism is somewhat limited, and that bees rely more on available food and less on body deposits. Interestingly, the non-metabolic characteristics also fluctuated over the year. The vitellogenin peak reached its maximum in the haemolymph in winter, which is probably associated with the immunoprotection of long-lived winter bees. The analysis of bee venom showed the maximal levels of vitellogenin in autumn; however, it is not entirely clear why this is the case. Finally, melittin levels showed strong fluctuations, suggesting that seasonal control was unlikely.
- Keywords
- Adipokinetic hormone, Metabolism, Seasonal fluctuations, Venom, Vitellogenin,
- MeSH
- Biomarkers metabolism MeSH
- Central Nervous System metabolism MeSH
- Hemolymph metabolism MeSH
- Insect Hormones metabolism MeSH
- Pyrrolidonecarboxylic Acid analogs & derivatives metabolism MeSH
- Melitten metabolism MeSH
- Oligopeptides metabolism MeSH
- Seasons * MeSH
- Digestive System enzymology MeSH
- Bee Venoms metabolism MeSH
- Bees physiology MeSH
- Vitellogenins metabolism MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- adipokinetic hormone MeSH Browser
- Biomarkers MeSH
- Insect Hormones MeSH
- Pyrrolidonecarboxylic Acid MeSH
- Melitten MeSH
- Oligopeptides MeSH
- Bee Venoms MeSH
- Vitellogenins MeSH
Bees originally developed their stinging apparatus and venom against members of their own species from other hives or against predatory insects. Nevertheless, the biological and biochemical response of arthropods to bee venom is not well studied. Thus, in this study, the physiological responses of a model insect species (American cockroach, Periplaneta americana) to honeybee venom were investigated. Bee venom toxins elicited severe stress (LD50 = 1.063 uL venom) resulting in a significant increase in adipokinetic hormones (AKHs) in the cockroach central nervous system and haemolymph. Venom treatment induced a large destruction of muscle cell ultrastructure, especially myofibrils and sarcomeres. Interestingly, co-application of venom with cockroach Peram-CAH-II AKH eliminated this effect. Envenomation modulated the levels of carbohydrates, lipids, and proteins in the haemolymph and the activity of digestive amylases, lipases, and proteases in the midgut. Bee venom significantly reduced vitellogenin levels in females. Dopamine and glutathione (GSH and GSSG) insignificantly increased after venom treatment. However, dopamine levels significantly increased after Peram-CAH-II application and after co-application with bee venom, while GSH and GSSG levels immediately increased after co-application. The results suggest a general reaction of the cockroach body to bee venom and at least a partial involvement of AKHs.
- Keywords
- American cockroach, adipokinetic hormone, dopamine, honey bee, melittin, metabolism, muscle ultrastructure, vitellogenin,
- MeSH
- Central Nervous System chemistry drug effects MeSH
- Hemolymph chemistry drug effects MeSH
- Insect Hormones pharmacology MeSH
- Pyrrolidonecarboxylic Acid analogs & derivatives pharmacology MeSH
- Oligopeptides pharmacology MeSH
- Periplaneta chemistry drug effects immunology MeSH
- Immunity, Innate * MeSH
- Bee Venoms adverse effects MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- adipokinetic hormone MeSH Browser
- Insect Hormones MeSH
- Pyrrolidonecarboxylic Acid MeSH
- Oligopeptides MeSH
- Bee Venoms 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.
- Keywords
- antimicrobial peptides, beta-alanyl-tyrosine, capillary zone electrophoresis, free-flow zone electrophoresis, peptide analysis, peptide purification,
- MeSH
- Anti-Infective Agents chemistry isolation & purification MeSH
- Dipeptides chemistry isolation & purification MeSH
- Electrophoresis methods MeSH
- Hemolymph chemistry MeSH
- Larva chemistry MeSH
- Sarcophagidae chemistry MeSH
- Chromatography, High Pressure Liquid MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- alanyltyrosine MeSH Browser
- Anti-Infective Agents MeSH
- Dipeptides MeSH
