Melanogenesis and melanin deposition are processes essential for the effective immune response of insects to various invaders. Phenoloxidase (PO), produced in specialized cells as an inactive precursor prophenoloxidase (proPO), is the key enzyme for melanin formation. The precursor is activated via limited proteolysis by a dedicated serine proteinase, which is the final element in the cascade of serine proteinases (SPs) that make up the PO system. Melanogenesis provides different cytotoxic molecules active in fighting infections, as well as melanin, which is important for sequestration of invaders. However, since the cytotoxic reactive compounds generated during melanization also pose a threat to host cells, strict control of the PO system is necessary for host self-protection. Different pathogens and parasites influence the PO system and melanization through various strategies, which allow them to survive and develop in the host insect body. In this review, we characterize “the lights and shadows” of PO system activation, indicating, on one hand, its advantages as an efficient and effective mechanism of the insect immune response and, on the other hand, the dangers for the insect host associated with the improper functioning of this system and selected strategies for regulating its activity by entomopathogenic organisms.

Antimicrobial peptides (AMPs) constitute a large and diverse group of molecules with antibacterial, antifungal, antiviral, antiprotozoan, and anticancer activity. In animals, they are key components of innate immunity involved in fighting against various pathogens. Proline-rich (Pr) AMPs are characterized by a high content of proline (and arginine) residues that can be organized into Pro-Arg-Pro motifs. Such peptides have been described in many invertebrates (annelids, crustaceans, insects, mollusks) and some vertebrates (mammals). The main objective of this review is to present the diversity of invertebrate PrAMPs, which are associated with the presence of cysteine-rich domains or whey acidic protein domains in the molecular structure, in addition to the presence of characteristic proline-rich regions. Moreover, PrAMPs can target intracellular structures in bacteria, e.g., 70S ribosomes and/or heat shock protein DnaK, leading to the inhibition of protein synthesis and accumulation of misfolded polypeptides in the cell. This unique mechanism of action makes it difficult for pathogens to acquire resistance to this type of molecule. Invertebrate PrAMPs have become the basis for the development of new synthetic analogues effective in combating pathogens. Due to their great diversity, new highly active molecules are still being searched for among PrAMPs from invertebrates.

The aim of this study was to investigate the changes in the level of oxidative stress and lysozyme-like and phenoloxidase (PO) activity under the influence of nosemosis. Honeybees were kept in natural (apiary) and artificial (laboratory) conditions. In this study, it was shown for the first time that honeybees kept in apiaries have higher levels and activity of the studied parameters than honeybees kept in the laboratory. The greatest difference was noted in the case of PO activity in 28-day-old infected honeybees in May, when the activity was 32.3 times higher in honeybees kept in the apiary than in the laboratory, suggesting that environmental conditions have a significant influence on the immune response of honeybees. Simultaneously, the apiary conditions resulted in higher level of oxidative stress, indicating lower effectiveness of antioxidative mechanisms. Additional nosemosis infection increased the level of oxidative stress as well as lysozyme and PO activities. In July, in 28-day-old infected honeybees kept in laboratory, the highest increase in PO activity (by 10.79 fold) was detected compared to healthy honeybees. This may indicate that infection causes a decrease in the effectiveness of primarily antioxidant mechanisms, whereas immune mechanisms are still activated during infection. Another interesting factor is the age of the honeybees. It was found that in the summer months (June, July) the lysozyme-like and PO activities increased with age, while in the case of oxidative stress the opposite trend was observed, suggesting better effectiveness of both immune and antioxidant mechanisms. Another important element is seasonality, which significantly affected only the lysozyme-like activity. It was found that in July in all the groups studied this activity was higher than in the other months. The results allow us to better understand the mechanisms of honeybee immunity, which are constantly being studied due to the complex social structure created by these insects. Our research emphasizes that honeybee immunity is dynamic and depends on a number of factors, such as environment, age, season or the presence of pathogens.

The honeybee is one of the most important pollinators in the world. The frequently observed poor health of honeybee colonies can be caused by various factors, e.g. environmental pollution, nutritional stress, and climate changes. Moreover, honeybees are constantly exposed to a wide spectrum of pathogens, such as parasites, bacteria, and viruses. We examined the occurrence of various diseases in different-aged worker honeybees from two colonies kept in natural and laboratory conditions during spring, summer, and autumn in Poland. The honeybees were examined by PCR to detect infection with selected pathogens: Nosema ceranae, N. apis, N. bombi, Acarapis woodi, trypanosomatids, and neogregarines (Mattesia or Apicystis species) and by RT-PCR to identify deformed wing virus (DWV), black queen cell virus (BQCV), and acute bee paralysis virus (ABPV). DWV and N. ceranae turned out to be the dominant pathogens. Trypanosomatids and BQCV were also found in several samples. We did not detect the presence of the other pathogens: N. apis, N. bombi, A. woodi, neogregarines, or ABPV. As shown in the present study, the dynamics and occurrence of pathogens are influenced by keeping conditions, honeybee age, and seasonality.

Aging is a multifactorial process that occurs in all living organisms, including bees. One of the factors accelerating this process is stress caused in bees by Varroa destructor. The research aim was to compare antioxidant system activities in different tissues and in different fat body segments (sternite, tergite 3 and 5) in workers aging naturally (physiologically) and prematurely (affected by V. destructor). The CAT, GPx, GST, and SOD activities in naturally aging workers were higher in all the tissues/fat body segments and age groups compared to prematurely aging workers. These antioxidant activities increased with age, reaching a maximum at 21 (in tergite 3 and sternite) or 28 days of age (in the hemolymph and tergite 5) in naturally aging workers, and then decreased in the oldest ones (at 35 days of age). In the prematurely aging workers, the antioxidant activities in the fat body decreased along with age. The highest activities were identified in the fat body of tergite 5, which may suggest its role in detoxification processes. Our results are a starting point for a better understanding of the mechanisms related to oxidative stress, aging, and their correlation with the health and lifespan of bees.

Recent years have witnessed a fall in the numbers of bee colonies worldwide. To a large extent, this is due to diseases affecting honeybees. One such disease is nosemosis, which is caused by fungi from the genus Nosema. The antibiotic fumagallin used to be administered to treat nosemosis, but as it was found capable of being transferred to bee products, this practice has ceased. We therefore focused on natural substances for treating this disease, among them, aqueous extracts of the nest carton produced by the jet-black ant (Lasius fuliginosus). We tested the influence of 0.1% and 1% concentrations of such extracts on the incidence of nosemosis in honeybees. The “birch carton 2” extract turned out to be the most effective inhibitor of this disease. The incidence of nosemosis following the administration of this extract, as manifested by the number of spores, fell ca 18-fold in comparison with the control. Moreover, the “birch carton 2” extract was not found to be toxic towards bees. GA